Экспрессия генов = Gene expression 5-02-001890-2

Citation preview

1 97

575 28.04 20

. . : . . . .

. . .– ISBN

.:

, 2000. – 000 .,

.

5-02-001890-2

, .

.

,

,

,

,

. ,

. .

, ,

,

,

-

-

. ,

,

. -97- -№ 123 Patrushev L.I. Gene Expression. – Moscow: Nauka, 2000. – 000 p., il. ISBN 5-02-001890-2

2

This monograph consisting of two parts presents a comprehensive view of the structure and functioning mechanisms of genes for prokaryotes and eukaryotes as well as the basic methods used for their studies. Its first part discusses the genome structure of prokaryotic and eukaryotic organisms, also covering the mechanisms of transcription, translation, replication, DNA repair and their regulation. The second part of the monograph deals with the principles of the techniques employed in the gene research. The book devotes particular attention to current genetic engineering tools. It embraces the key aspects of recent developments of molecular genetics studies on site-directed mutagenesis, protein engineering, antisense RNA, ribozymes and deoxyribozymes, transgenosis and gene therapy, DNA diagnostics and genotyping human genome. The book will act as authoritative references for researchers, postgraduates, and students specializing in chemistry and biology. ISBN 5-02-001890-2 ©

"

", 2000

3

................................................................................. 9 ..................................................................................... 12

I. ................................. 17 18 1.

................................................................................................ 25

1.1.

.......................................................................... 28

1.2.

.............................................................................. 30

1.2.1. 1.2.2. 1.2.3. 1.2.4. 1.3.

.................................................................................... 31 ...................................................... 32 .......................................................................... 35 ............ 37 ................................................................................ 40

1.3.1. 1.3.2. 1.3.3.

........ 41 ............................................................................................ 45 ................................................... 55

2. ...................................................................... 60 2.1.

................................................................................... 61

2.1.1. 2.1.2. 2.1.3. 2.1.4. 2.2.

-

-

.................................................... 62 ( ) ......................................... 72 .......................................................................... 80 .................................................. 112

................................................................................................ 124 2.2.1. 2.2.2. 2.2.3. 2.2.4.

2.3.

............................................................ 125 ............................................................. 131 ................................ 143 , II ............................................................................ 163 ......................... 168 -

-

2.3.1. 2.3.2. 2.3.3. 2.3.4. 2.3.5. 2.4.

................................................... 169 ........................................................................................... 171 ............................. 173 .............................................................. 176 ...................................................... 178 .................................... 180

2.4.1.

......................................................................................... 180

4 2.4.2. 2.4.3. 2.5.

................................................................. 187 ...................... 197 ................................................................. 200

,

2.5.1. 2.5.2. 2.5.3. 2.5.4. 2.5.5. 2.5.6.

......... 201 .... 202 ................................................... 212 ................................................................. 213 ........................................................... 215 ............................................................ 219

3.

............ 223

3.1. .................................................................................... 225 3.1.1. 3.1.2. 3.2.

............................ 225 ....... 229 ....................................................................................... 238

3.2.1. 3.2.2. 3.2.3. 3.2.4. 3.2.5. 3.2.6. 3.3.

.............................. 241 ......................... 259 ......................... 297 .......................................................................................... 304 ...................................................................................... 318 ........................... 319 ............. 330

3.3.1. 3.3.2. 3.3.3. 3.4.

, ............................................................... 330 .............................. 343 .................................................. 353 .............. 358

3.4.1. 3.4.2. 3.4.3. 3.5.

................................................. 358 ................... 367 ............................................... 370 ........... 371

,

3.6.

................. 373

3.6.1. .......................................................................................... 373 3.6.2. ........................................................................................ 376 3.6.3.

-

3.6.4. 3.6.5.

.................................................................... 377 ............................................................................ 381 ...................... 386

4.

........... 390

4.1.

............................................................................. 390

5 4.1.1. 4.1.2. 4.1.3. 4.2.

, E. coli

........................................ 391 T4 ........................... 394 400 ......................................................... 403

4.2.1. 4.2.2. 4.3.

E. coli .................... 405 ColE1........................................ 414 .......................... 421

4.3.1. 4.3.2. 4.3.3. 4.3.4.

..................................................... 422 .................................................................... 425 ... 427 ............. 429 Щ

5. 5.1.

................................. 432 ........................................................................................... 433

5.1.1. 5.1.2. 5.1.3. 5.1.4. 5.1.5. 5.1.6. 5.2.

................................................................................. 435 .............................................................. 449 ...................................................................... 453 ................................................................................. 456 ....................................................................... 460 ......................................... 464 ............................................................................... 465

SOS-

5.2.1. 5.2.2. 5.2.3. 5.2.4. 5.2.5. 5.3.

................. 466 .............................. 468 ............................ 481 ............................. 483 (ADP) ..... 486 ..................................................................... 490

5.3.1. .................................................................................................... 491 5.3.2. ............................................................ 495 .......................................... 500

5.3.3. 5.3.4. 5.3.5.

............................................................................... 508 ..................................................... 516

6.

............................................ 522

II. .......................................................................................................... 531 7.

............................................. 532

7.1. 7.1.1. 7.1.2. 7.1.3. 7.1.4.

, -

-

... 539

......................................................... 539 .......................................................................... 548 .................................... 549 ............................................................................ 556

6 7.2.

........................................................................................... 558 ...................................................................... 559 λ .............................................................. 563 ........................................................................ 568 YAC, BAC PAC ........................................ 570 ( ) ....................... 574

7.2.1. 7.2.2. 7.2.3. 7.2.4. 7.2.5. 7.2.6. 7.2.7. 7.3.

................................................................... 577 ......... 593 ............................................................................. 596

7.3.1. 7.3.2. 7.3.3. 7.4.

.............................................................. 596 ...................................... 599 .................................................. 600 , ............................................ 602

7.4.1. 7.4.2. 7.4.3. 7.4.4. 7.4.5. 7.4.6. 7.5.

( ( ,

7.5.1. 7.5.2. 7.5.3. 7.6. 7.6.1. 7.6.2. 7.6.3. 7.6.4. 7.7.

............................................................ 615 .............................................................. 618 ......................................................................... 619 ................ 621 ............................................. 621 " ............................................. 622 ........................................................... 624 ..................................................................................... 625 ............................................. 626

" S1-

8.

... 629

8.1. 8.1.1. 8.1.2. 8.1.3.

............................ 630 ........................................................ 630 ..................................................................... 633 ..................................................................... 635 ....... 642 ..................................................................... 646

8.1.4. 8.2. 8.2.1. 8.2.2. 8.2.3. 8.2.4. 8.2.5. 8.3.

( CHO)....................... 604 Sp2/0) ...................................... 607 MEL) .......................................... 608 ( COS) ................. 610 ......................... 611 .. 612 ........................ 613

-

................................................... 646 ........................................... 655 ......................................................................... 656 ......................................... 662 ............................................ 666 .................................................................. 670

7 9.

, .......................................................................................................... 679

9.1.

................................ 679

9.1.1. 9.1.2. 9.1.3. 9.1.4.

...................................... 680 ............................................. 682 ................................................... 687 : .................................... 690 ................................................... 692

9.2. 9.2.1. 9.2.2. 9.2.3. 9.2.4.

............................................................................... 692 ........................................................................ 694 ......................................... 698 , ......................................... 704 ............................................................................ 708 ........................................................................................ 710

9.2.5. 9.3. 9.4.

.................................................. 713

9.4.1. 9.4.2.

-

10.

...................................... 714 ... 717 ............................... 722

10.1. ....................................................................................................... 722 10.2.

................................................................ 725

10.3.

................................... 726

10.3.1. 10.3.2.

............................ 727 .................................................................. 728

10.3.3. ............................................ 729 10.3.4. .................................................................................... 732 .................................................................. 735

10.4. 10.5.

....................................................................................................... 737 10.5.1. 10.5.2. 10.5.3.

-

..................... 738 ......... 743

.............................................................................. 746 ............. 750 ..................... 752

10.5.4. 10.5.5. 10.5.6.

, ............................................................................... 753

11.

-

-

.............................. 755

8 11.1.

................................................................................ 759

11.1.1. 11.1.2. 11.2. 11.2.1. 11.2.2. 11.3. 11.3.1. 11.3.2. 11.3.3.

.................. 759 ............................................................ 761 ........................................................................... 774

-

........................................... 775 : ............................................................ 778 .............................................. 786 .......................................... 787 ..................... 790 ..................................................... 792

12. ...................................................................... 796 12.1. 12.1.1. 12.1.2. 12.1.3. 12.1.4. 12.2.

.......... 796 ................................................... 797 ..................................... 802 ...................................... 804 ................................... 807 ...................................................................................... 809

12.3.

.................................... 810 .................................................................................................. 813 .................................................................. 818

9

.

. 1950–1960-

–

,

,

– . –

coup

,

de

.

force,

,

,

Biology",

"Journal of Molecular

.

,

"Nature New Biology" “

: «

”, ,

,

,

,

».

,

: , . , :“

,

,–

, 1980

1970–1980-

”.

.

,

( ),

,

, .

, ,

10 .

,

,

.

.

–

.

,

. .

, . .

,

, ,

. ,

,

.

, . -

,

(

)

–

–

-

. . , . , .

. –

. .

,

,

.

, , ,

. ,

, ,

– ,

.

11 – . .

, ,

,

,

, – .

. 1999 .

12

-

, … ,

,

.

,

,

1970.

,

.

.

,

.

,

, 25

,

. -

,

.

,

, ,

,

-

.

–

,

. "

" ,

, .

,

70-

XX .

, . ,

,

,

, . , , ,

. -

13 , . .

. .

.

(

),

,

, ,

-

. .

–

, , –

-

, . , ,

,

.

,

,

, .

. , . , .

, , -

,

.

,

,

-

( .

,

-

)

14 , 2020

. .

, .

,

.

,

.

,

.

.

,

, ,

. . , . .

, ,

, .

. "

"

. ,

.

,

15 , "

", .

, .

.

, .

. ,

,

, .

,

,

,

. ,

,

. .

,

, ,

.

,

, ,

,

,

.

. . .

. . .

, . .

,

. .

16

. .

, .

,

, ,

.

, , -

. .

,

. , ,

,

,

. . .

. .

,

. .

,

. . . . . , .

. .

, .

, ,

.

,

, ,

,

,

, .

.

, . , ,

,

,

. . .

17

I.

18

. ,

,

,

. .

(1998

.) XX

.

,

,

.

, , , . , , status quo

. ,

, ,

, .

,

,

,

, . ,

(

), "

"

,

.

, , .

.

19

. .

, .

:

. ,

, ,

–

.

. ,

. ,

. ,

.

, .

–

.

.

,

, .

,

, .

,

, ,

(

)

, .

,

,

.

,

, , –

,

.

,

, , .

,

20

,

,

. ,

, , . "Omnia mea mecum porto!1" – .

. ,

,

, ,

–

. . ,

,

. ,

.

. ,

, ,

.

.

,

,

,

. ( ),

,

. .

, (

)

. , ,

.

1

В

(лат.)

21 , .

(

)

,

,

(

)

–

.

–

– . .

(

). , –

,

. ,

.

.

. 14

~10 –10

,

15

100

.

. . ,

. ,

, .

.

(

)

,

,

. ,

. . .

22

.

,

–

. ,

.

,

, . , ,

. , , .

. ,

, ,

, . :

-

,

,

. , ,

, . .

– –

, .

. ,

. ,

,

23 (

)

(

)

. .

– .

,

,

,

, ,

.

(

) . ,

,

, .

. , , .

,

,

(

,

) .

– , .

. –

.

,

24 .

,

,

,

, ,

.

,

. , ,

, . , ,

,

,

, .

25

1. "

"

.

1920 .

, . , ,

. .

,

, ,

,

, .

("

")

, ,

.

.

. . , ,

– .

,

, (

)

,

. , ( .

)

,

, .

.

, ,

,

-

,

26 .

-

,

,

, , . . I.1, .

. I.1

, . . 1,0·104 1,6·106 2,0·106 4,7·107 2,3·109 1,6·109 1,4·109 2,7·109 3,6·1010 1,5·109 1,2·109 2,6·109 3,0·109 1,6·1010 2,7·1010 Lilium longiflorum

1,8·1011

27

.

1978 . .

-

, (∼3%

"

".

(

.

).

5.3), -

,

,

, .

, . : .

,

,

, .

:

(Archaebacteria),

(Shizomycophyta)

(Cyanophyta).

,

,

-

,

,

,

. , :

. ( (Planta)

) (Mesokaryotes),

(Fungi),

(Animalia). ,

, ,

, . ,

,

,

,

28 . 1.1.

.

, ,

,

.

(

) ,

, .

, (

)

,

.

(

) –

.

,

,

,

, .

.

, . ,

,

(>60 . . .) ,

.

(

) (

,

).

, .

,

, ,

.

,

29

, .

. ,

, in vivo. , , . (

–

,

( ( (

)

)

)

.

,

)

, ,

.

.

, ,

.

, , . 1%

, ,

Sacharomyces cerevisiae .

20% (

)

1

10%

.

,

, ,

. , .

,

30 , ,

.

Amphiuma

~30

,

,

28 (46

,

, ). .

, .

,

Lysandra nivescens

380–382

,

, .

-

, ,

. .

.

,

,

,

, . 1.2. , ( ,

)

,

–

,

. , .

,

. , ,

, .

,

,

. ,

,

.

31 , .

,

,

,

,

,

,

, , , . 1.2.1. .

-

,"

–

,

,

(

)

; -

-

; (

),

"

", 2

-

". , .

-

. .

, ,

. – , , .

2

-

.

.

.:

, 1972.

. 15.

32 ,

( ).

,

,

(

). .

.

-

, ,

, ,

. ,

, .

(

,

),

.

, .

,

, (

),

(

).

-

. -

, ,

.

,

-

,

, , .

,

.

1.2.2. ,

, ,

33

. I.1.

E. coli

–

,

(

.1 2 .

). ,

2– 3 ; .

, – .

.

,

34 .

-

, (

∼4,6

.

E. coli

. .) . , . , ,

,

. -

. ,

,

-

.

,

,

,

, .

, ,

(

. I.1, ).

. -

, ,

-

HU

I . ,

.

,

, . . I.1, .

.

.

,

, , .

35 . , , .

, ,

.

,

-

,

, -

HU, H-NS

, IHF,

,

. "

" (

)

. LP-

(low

protein chromatin), .

LP-

,

,

(

).

,

,

.

. : genitalium

E. coli

Haemophilus influenzae. ∼4,6

Bacillus subtilis

1997 .

4,2

. .

,

. 1.2.3.

.

Mycoplasma

36 Archeabacteria

, . ,

. Methanococcus jannaschii, 1996 ., .

.

,

,

, 200

. M. jannaschii :

– CO2, NH3

. .

M. jannaschii , 1700, 58

16 . . .

.

,

-

GC31%.

∼1700

: ,

1000 . . -

M. jannaschii .

.

,

.

M. jannaschii

M. jannaschii

,

, –

,

, ,

. (

)

.

–

,

. ,

,

M. jannaschii .

37 .

,

,

-

-

. , -

M. jannaschii ,

, , ,

-

.

M. jannaschii

, ,

"

"

.

, ,

, . M. jannaschii

,

,

-

ε.

Pol III,

-

,

M. jannaschii. :

,

,

,

, ,

.

M. jannaschii,

,

. , -

,

,

-

.

, .

M. jannaschii .

, , . 1.2.4. , ,

38 , . , ,

,

,

. .

1920-

.

(1995 .) (Mycoplasma genitalium

Haemophilus influenzae) .

. .

. . , , .

,

M. genitalium

H. influenzae

-

1,5

.

,

, , , . ∼580 H. influenzae (~1830

. . .,

. . .)

,

M. genitalium,

469

,

1703

M. genitalium

.

240

,

,

H. influenzae. 22

, , .

6

,

, . ,

256

,

39 . ,

256

,

:

; ; ;

, ;

,

,

;

,

, ;

;

,

, ;

;

,

;

de

novo

;

(

).

, . Bacillus subtilis, . ,

318 . . ., 562 ,

. . . M. genitalium.

. .

, .

,

,

M.genitalium

H. influenzae, (∼ 250),

.

, ,

40 . 1.3. ,

(

),

,

–

,

.

,

, ,

.

-

.

–

.

.

, . . .

,

. – ,

1

(

168

∼6

).

6·109 . . (

,

, (

)

.

. I.1).

, ,

, .

, , .

,

41 ,

.

, . 1.3.1.

. , , 15–98%

, .

, ,

:

.

,

,

, . (

–

)

, , -

. .

(1977 .),

,

-

,

, (exon shuffling). ,

(

),

. , , ,

,

42 "

. " (intron late). . .

. .

(1978 .)

"

".

-

.

. . 105

, ,

10–104

,

. ,

1–20

. .,

.

.

,

(

,

). ,

,

, .

. .

. (1974 .)

–

,

, CsCl. ,

: ) ;

; )

)

;

)

( );

)

– ; )

; (

)

)

; (

)

) .

43 5–50% .

-(

)

(

1

4 . . . .

ó

)

, .

VNTR

(variable number of tandem repeats). – , .

, ,

(medium reiterated frequency repeats – MERs), : SINE (short interspersed elements) –

LINE

(long interspersed elements) – SINE-

. 90–400

. .,

LINE-

7 . . . SINE ∼300

Alu-

, ~106

. . Alu4

. . .,

~5%

.

,

B1, . LINE,

( , ), (long terminal repeats – LTR), (

.

LINE-

7.2.7). LINE-1-

. ORF-1

LINE-1ORF-2,

,

44 ,

. ORF , (SDR).

LINE-1

–

5’.

LINE-1

(

)

A 600

F,

.

. .

A- (

F)

. , SINE-

LINE-

. .

,

, ,

20–30%. , , ,

.

-

, ,

.

. LINEORF,

-

, ,

,

– , . 100 000

MaLR-

2–3 . . .,

LTR, .

, .

"

"

,

45

.

,

,

"

"

2–3 ,

.

, "

"

,

,

"

",

.

"

,

,

"

,

, , . :

, -

?

,

.

,

C,

,

-

. "

"

5.3. 1.3.2. , .

, , .

~1:1,

.

46 , H1, H2A, H2B, H3

,

~220 (H1) . H2A

102 (H4) Lys,

H1

H2B

Lys,

H3

Arg.

H4

(

H4) . H1 ,

.

H1.1–H1.5, H1

o

H1t.

. I.2.

-

– MAR/SAR-

; ;

–

–"

",

"

"

. ∼10

H4.

,

, 1

.

, ,

,

~4 ~4

.

. ,

,

,

. . , .

∼104

.

47 . , . .

.

.

, .

.

, . .

.

,

,

, . in vitro , . ;

: 1) 2)

,

;

3)

,

-

. .

( )

10

,

.

,

,

.

,

, 10–20

.

48 : H2a, H2b, H3

H4,

H1. : , – .

.

, (

10

2

. .)

3

4,

2b.

,

B-

. .)

, –

(146

11

– 5,5

.

,

ó

,

. .

ó ,

,

200 (±15)

. .

.

N, (arm).

-

, -

–

, ,

ó

α, . , , (

. I.2).

N-

H3 ,

– .

S. cerevisiae N-

.

H4

,

49 . 3

4

( 3)2( 4)2,

2

2

–

. ,

,

2 / 2 , 3/ 4.

.

. (1997

.)

.

,

27–28 . ., 140 , 4 . .

B-

ó 0,25–0,35

. ./

, – 10,5

. ./

: ,

10,2 . ./

(

-

). ,

ó

. N-

,

-

, .

, N-

3 ,

ó 4

.

,

121

. .

H3. N-

H3 ,

,

N-

H2A .

, .

H2B

50 , 15

10–

40 . .

.

I.2

(

)

N-

3

135

-

41

25

Ac-K9, P-S10, Ac-K-14, Ac-K18, Ac-K23, Ac-K27

H4

102

32

–

P-S1, Ac-K5, Ac-K8, Ac-K12, Ac-K16, Ac-K20

H2A

129

24

16

P-S1, Ac-K5, Ac-K9

H2B

125

30

23

Ac-K12, Ac-K15, Ac-K20, Ac-K24, P-S32, P-S36

H1

213

36

92

S-

: P-S145, P-S173, P-S180 : P-S16, P-T17, P-T136,

P-S145, P-S173, P-S180 . Ac –

, P –

Lys, Arg, Thr

Ser

; K, R, T

S –

. .

"

"

in vitro 1

. 1, ,

, 30

,

.

, in vivo. 25~30

,

51

(∼100

NaCl),

. . 30 .

,

,

, "

(10

. .

,

" (super-beads).

.

∼6

)

,

, 10

~6

,

–

40

.

,

,

. . ,

, . 1, ó

30

. 1,

1,

1 ,

1 ,

5, . 1,

,

ó . 1

, ,

ó ,

.

1,

, . , .

,

, ,

1 ,

52 . , . -

. , ,

, ,

100

50–

. . .

,

(

)

.

. I.2).

. , , ,

~5

. . . (

, , (

) –

,

.

,

, MAR (Matrix Associated Region)

Associated Region)

SAR (Scaffold

MAR/SAR-

. MAR/SAR. , ,

-

II,

, .

(∼10 . . .),

"

,

"

, ,

, . I.2).

( , MAR/SAR-

.

, ,

. MAR/SAR-

53 . 200–350 . .,

(dT).

(dA)⋅ -

AATAAAT/CAAA

-

:

-

–

– TTA/TTT/ATTT/CAAA,

MAR/SAR-

,

. –

ATATTT

AATATTTT,

MAR/SAR, . MAR/SAR-

,

. .

MAR/SAR-

(1995 .):

300–1000 . .; -

AT-

; MAR/SAR-

, ,

; MAR/SAR-

3–112

. . ., , -

; MAR/SAR.

,

; MAR/SAR,

Z-

I. ,

MAR/SAR-

H-

,

54

.

,

, , , .

MAR/SAR( ,

.

), 3.2.4. -

, MAR/SAR-

. ,

. . .

. .

(1998

.)

,

,

SAR-

,

,

, .

,

MAR/SAR,

,

,

,

. .

. , (high mobility group – HMG). . ~10 : 14/17, 1/2

,

HMG.

I/Y.

HMG 14/17 10–12

, . N-

55 , –

.

-

ó

HMG 14/17, . HMG-

H1 . In vivo ~10% HMG 14/17. HMG 1/2 25–30

. ,

,

, . ,

(

)

.

1/2

,

HMG

in vitro, ,

. HMG I/Y

-

,

1.

,

1 in vivo (

.

2.1

4.1).

HMG, . ,

, ,

.

-

, . 1.3.3.

-

, , ,

, .

56 , ( -

). I

II.

-

I,

,

. I.3.

II

. 5'-

. VM26, mAMSA, II,

– ;L– .

. ,

,

-

II

ATP( Tyr, .

. I.3).

57 5'-

3'-

.

. I

-

Tyr

,

-

II , .

,

,

-

I – IA

,

IB,

, , .

-

IA

, ,

,

-

III. -

IA

, Mg2+

, 5'-

é

.

-

IA

II

,

.

,

IB

-

, , 3'-

.

IB

( ). -

IB/III

(camptothecin), .

I–

, .

. -

-

II

58 . , (

,

). -

II

I. -

II

, .

-

II

,

. -

II ,

.

,

.

-

II

.

-

. ,

-

II ,

MAR/SAR-

.

II

, .

,

in vivo

, : ,

, MAR/SAR-

.

– -

,

II

59 .

,

-

II

, , , .

β)

(α

,

-

,

.

, ,

.

, . ,

-

,

.

. , ,

. ,

(

MAR/SAR)

, .

, .

,

-

,

, ,

(

.

5.3).

, (

) .

60

2.

, , , .

↔

:

.

→

,

,

. . , ,

(

)

(

– messenger RNA, mRNA),

. , . ,

,

(

), .

( ,

), ,

,

.

,

. "

"

, ,

,

. .

,

,

61

.

, -

: , ,

. .

, (

).

. . 2.1. , , (ATP, GTP, CTP '

UTP)

'

3 –5 . ,

, -

-

, –

,

–

–

. , -

.

62

. 2.1.1.

-

.

, ,

–

,

-

SP6

T7.

, .

-

, , .

, .

,

-

-

.

,

,

-

in vivo , . -

E. coli. -

E. coli. .

(

165

), β- (155

α- (35

),

)

ββ‘αα,

(σ70)).

70

σ- (

(enzyme), )

: β‘-

(

-

E. coli, ,

(

,

.

).

σ-

,

,

,

63 -

–

,

,

.

σ70-

,

σ . 70

(

)

E. coli.

,

(

54

gln

,

nif –

σ70

)

σ70

-

Eσ54.

σ-

,

.

-

-

,

σ54

, β-

α.

β‘-

.

,

,

. , ,

α-

,

,

.

, (

),

, ,

–

, ,

.

. – – . ( )

.

. ,

,

,

64 σ-

, –

-

. -

I 18S , (

28S

-

;

.

-

(

),

-

II

III

), 5S

. , (120–220

)

5–13

(10–100

)

. -

.

,

β-

,

β’-

, .

α-

, -

, , I

II,

,

-

III.

-

,

, -

, ,

, -

. -

I

(Pol I). ,

: ,

.

.

65

. I.4. -

I( ) ( )

66 – , , -

.

Iα-Iδ

-

I.

, Mg2+).

( ,

.

– , . , .

↔ ,

(RPA194

RPA116)

, , → – 5′-

. , Eσ70,

I , . tss –

.

.

in vitro . , Pol I

. I.4,

,

'-

E. coli.

-

I

,

11

.

Pol I 194

, ), 15–60

.

Pol I 53

Pol I Pol I*)

116

(

14

53),

-

,

PAF53 (polymerase associated factor Pol I

RPA49

,

-

,

RPA49

.

RPA35.5 (

67 poly[d(A-T)],

.

, (

.

). Pol I

,

,

,

,

Pol I

, .

Pol I . TIF-IB (Pol I-specific transcription initiation factor B), ,

D, (

– rSL1

Pol I

).

hSL1,

X. laevis – Rib 1.

TIF-IB/SL1 Pol I .

TIF-IB/SL1

,

TBP, -

(

. .

2.1.3.) 110, 63

48

TBP-

TAFI, TBP,

,

. TIF-IB/SL1

UBF ( .

TAFI48 .

),

TAFI63

TAFI110

TAFI TAFII,

,

Pol II. TBP

TBP

TAFI

TAFII (

), .

TAFI48

TBP

-

,

TATAPol II-

, ,

–

,

Pol II. ,

Pol I, UBF (upstream

binding factor)

. UBF ,

-

68 HMG-

(high mobility group domain) – .

80

-

, α-

HMGL,

,

-

L.

UBF 97

– UBF1 95

UBF2

,

. UBF1

HMG-

,

N-

C,

.

HMG-

UBF

,

UBF

(

).

,

HMG-

,

UBF.

,

– Pol I

TIF-IB/SL1, .

-

UBF

Ser .

,

HMG-

,

.

UBF, . CPBF (core promoter binding factor), , (

.

2.1.2.). CPBF, USF1

Pol I,

USF2 USF1

Pol I,

44

39

.

USF2 USF1/USF2

,

in vitro.

CPBF

Pol I in vivo.

TIF-IA –

Pol I, . ,

,

. TIF-IA

69

. 70

TIF-IA

. TIF-IA

70–80

. , , .

,

TIF-IC

Pol I.

, .

, .

TIF-IC TFIIF (RAP30/74) Pol II (

-

II

(Pol II).

.

).

Pol II

10

,

. (GTFs – general transcription factors). Pol II .

. .

, .

,

,

Pol II

14 . I.3.

,

70 I.3 -

Pol II

II

-

,

,

TFIIB

Pol II

TFIIF

TBP

,

Pol II, Pol II,

TFIIH

SRB/

-

ATP

,

,

-

CTD-

SRB2, SRB5

, , TBP, SRB/

GAL11/SPT13

, , SRB/

SUG1

SRB/

SRB9, SRB10, SRB11

,

SRB/

SRB4, SRB6, SRB7, SRB8,

,

CTD-

, Pol II

71 , ,

, SRB-

(suppressor of RNA polymerase B). Pol II

. SRB, -

CTD-

Pol II ( -

II). SRB.

,

SRB-

in vivo. SRB,

CTD-

Pol II,

-

,

. Pol II

GAL4–VP16, Pol II. ,

Pol II

,

.

SRB-

,

,

CTD,

∼10

, TFIIF, GAL11/SPT13, SUG1 -

Pol II, ,

. . Pol II

(

.

3.2.2).

-

,

,

. ,

,

,

.

,

-

, .

,

T-

,

72 , -

-

,

,

T7. -

, ,

-

-

-

,

,

-

,

,

.

,

,

,

-

, .

, ,

-

. ,

(

)

-

,

T7, . .

.

.

,

, -

, .

,

,

,

-

,

(β –

)

,

,

,

. 2.1.2.

( -

) in vivo

,

, –

.

, ,

, . ,

"

. "

"

"

73 "

",

.

,

,

, . , .

,

, -

,

. . (

)

,

, -

, , . .

1964 .

.

lac-

,

.

.

E. coli. , -

,

.

,

: 1) ; 2)

-

-,

,

; 3)

, , .

,

(

)

. , ,

74 –

.

(Eσ70)

-

. I.4, . :

-35,

–30 –10,

–7

–35,

–10 (

"

"

, ,

+1). σ70,

σ4.2

.

17

σ2.4.

. .

.

,

, ,

–

UP-

-

, P1

α-

–52

rrnB,

.

-

.

TG-

(

–

–15

–14),

. –10

–35

,

,

, . , ,

in vivo, –43 ( –1

USR – upstream region), +20 (

DSR – downstream region). -

(

.

,

). .

, ,

.

75

3.1. . , (

)

. -

Eσ. -

II.

,

-

II, . ,

ó

, (

.

. I.4, ). :

TATA-

,

25–30

(

– TATAa/tAa/t),

(Inr), (

– PyPyA+1NT/APyPy). , , Py –

,N–

.

, (

).

TATA-

.

-

II .

. ,

(

50

), (

), 60 . . .

76 , .

. I.5.

II

1–

n

– ,

1– n – , – , +1 –

,

-

–

II . I.5.

–

– . CA, +1.

, ,

TATA-

.

10–15 . ., .

,

,

, (

).

.

, , "

"

. -

(

.

. I.4, ).

1-

III.

,

III, 5S

,

,

77 (internal control region – ICR). ICR, TFIIIA,

+50

+64.

TFIIIA–ICR

TFIIIA

(

.

1

TFIIIC,

2.1.3

3.2).

,

2,

,

. . A B.

+10 30–60

+20,

B

. .

3’-

. TFIIIC,

.

,

,

TFIIIC–

, –25

+75.

. ,

U6-

,

, . -

–30

PSE (proximal sequence element) –60.

-

,

, PSE

. -

II

-

,

PSE,

. ,

.

PSESNAP -

PTF. (TBP).

,

78 TFIIIB, -

. PSE-

,

,

PSE-

,

TFIIIB.

U6-

,

III,

-

U2-

,

II,

-

U2-

. ,

U6 Pol II-

.

-

, II

Pol III-

,

III

,

-

, -

.

I.

,

,

,

, ,

,

. ( ). ,

,

,

/ .

,

,

.

, .

-

ό

I

(CPE – core promoter element), –30

+1 (

.

–75

–50,

. I.4, ).

TBP-

SL1 (

),

,

,

(TBP

associated factor – TAF),

(

. TBP

I.

2.1.3) ,

-

79 I

TATA-

, TAFI63

.

TAFI110,

SL1,

-

. 7.6.4)

(

,

.

SL1 -

I.

SL1 -

UBF,

UCE (upstream control element), –90 SL1

–150.

UBF

-

I .

, Pol I,

, . CPE

,

.

UCE -

I.

,

, UCE

,

UBF .

, -

II

TFIIB. BRF -

-

I

.

,

.

,

– CPBF,

ó

binding factor),

III.

1, – E1BF (enhancer 1Pol I.

80 2.1.3. 4 :

1)

; 2)

; 3)

; 4)

.

, ,

,

,

.

-

, .

, – . ,

, .

.

, ; . , . I.6. . ,

II

,

.

,

, . ,

,

,

(

. ) .

-

81 ,

.

,

in vivo, in vitro,

.

. I.6.

-

. .

,

-

, , . , ,

E. coli,

α-

β-

σ.

-

E. coli (

.

, ,

-

σ-

)

82

−35-

α-

. .

~50

.

. , -

.

-

E. coli , -

II

. -

II. (

)

.

,

Pol II ,

.

, , .

(

. I.4). , , .

.

,

.

83 I.4

(GTF)

-

II

(GTF) (

TFIIA

)

,

37 (α) TBP

19 (β) 13 (γ) TFIIB

,

TAF

TATA-

,

35

Pol II/TFIIF

, TFIID

-

38 (TBP)

, -

,

250 (TAFII250) 150 (TAFII150)

3’-

135 (TAFII135) 95 (TAFII95) 80 (TAFII80)

H4

55 (TAFII55) 31 (TAFII31)

H3

28 (TAFII28) 20 (TAFII20)

H2B

HMG-

,

84 1.4 (

)

(GTF) (

TFIIE

)

56

Pol II (

34

– RAP),

Zn-

, , TFIIH

TFIIF

58 (RAP74)

,

σ-

in vivo

-

E. coli,

26 (RAP30)

TFIIH

89 (ERCC3)

3’→5’-

,

;

80 (ERCC2)

5’→3’-

,

;

62 (p62) 44 (hSSL1)

Zn-

40 (cdk7)

,

-

Pol II (CTDcdk7-

37 (

H)

(CAK)

Zn-

,

Zn-

;

hSSL1 34 (p34) CAK 32 (MAT-1) TFII I

120

INRTBP

(

),

),

85

. I.7.

-

II

–

;

–

, ,

. (

,

, TAF-

–

(TAB-associated factors)) ,

86 ,

. 3.2.2. TATATFIID (transcription factor II D),

,

TATA-

(TBP), (

.

. I.7, ).

TBP

-

I

III,

.

TBP

TATA, .

TATA-

TFIID TFIIA

TFIIB

.

-

TFIIB. II

II TFIIE, TFIIF,

TFIIH

TFIIJ, (

.

. I.7, ).

,

,

, –

. ,

. .

,

. TFIIB,

TFIIE

σ70-

TFIIF -

E. coli. .

-

, .

,

β-

β‘-

E. coli , -

87 . −10

+3

.

, (

7–8

)

, ,

(

-

). . ,

σ-

-

E. coli ,

,

.

-

.

. -

I

III),

-

E. coli ( II

(

52

) CTD (carboxy terminal domain), . , Ser

CTD

Thr.

,

in vitro CTD, TFIIH.

, cdk7,

.

in vivo

,

CTDII

-

,

, . " (promoter clearance (escape)) (

" .

. I.7, ).

,

88 ,

-

II,

ATP-

. , ,

, .

,

(

)

TFIIH (

.

. I.4,

5.2.2).

. .

. -

σ-

E. coli:

, ,

-

. -

, .

, , ,

,

–

(

-

(

CTD-

II).

). .

-

: , . . -

, .

89

. I.8.

E. coli ( )

: .

– I

II, ,

( ) I

II – ,D U– , – 3′-

–

.

:

(1), (2) (3). I (

II – ; III –

.)

; IV – (

-

;

.) ,

90 –

-

I; –D – -

3′( . .)

, (

∼ 17

,

2000 . . .

),

,

. -

, –

–

. ,

,

-

,

+4 . , . -

, , . ,

(walking). , . . .

,

,

-

,

, -

-

.

in vitro

200–400

/ .

– 50–100

/ ,

in

-

vitro

II 5–10

in vivo

/ .

91 ∼20–30

/ . ,

-

,

. 10-3–

10-5.

,

,

,

3’→5’-

-

5’→3’-

.

3’(

, .

).

, .

,

.

, , . .

,

,

-

-

(

.

. I.8). ,

-

,

.

,

,

,

. ,

, (transcription bubble),

-

. , (

)

, .

,

, -

.

, 3’-

-

92

.

,

. ,

,

2–12 . .

, . , , -

, .

-

,

,

-

. .

, .

-

E. coli

, ,

–

(

)

,

. . .

-

, (pause),

(arrest),

.

, .

,

,

, .

,

-

,

, , -

,

– .

93

. ,

, . ,

.

, , . , – . , -

. , ; ,

. -

, ,

90%. ,

100%. 7

,

. ,

-

, , , . -

, .

é

94 -

,

-

.

I, II

III

,

, (

.

3.2).

é

,

-

,

. ,

.

in vitro .

,

, . , ,

,

.

é , .

in vivo.

, ,

,

, .

-

, . , . ,

,

95 . ,

,

.

,

.

-

, .

,

, .

,

,

, .

,

,

. ,

IMP

GMP, ,

,

, .

,

, .

, -

,

.

3'-

.

, . ,

.

,

,

17 ,

.

, . -

.

E. coli -

.

, . ,

96 .

, ,

,

.

,

,

-

. ,

GMP

IMP,

, . ,

, . , .

.

,

,

. I

,

-

II

,

.

-

, ,

-

. .

-

II

. , .

, ,

.

, . . ,

. , ,

97 ,

.

,

-

, -

,

.

-

. ,

, .

,

,

. .

Z-

,

, .

,

-

E. coli .

7-

1–5

.

24

,

.

, .

, 3’-

-

,

,

: 3’-

,

3’-

-

. ,

.

-

,

-

-

E. coli, 3’-

.

5’,

3’-

.

-

-

,

, . ,

,

5’-

,

98 5'-

.

E. coli GreA

GreB,

TFIIS ,

.

, 3’1

,

17

.

.

, ,

. , ,

,

,

. ,

, ,

5’-

,

,

. -

, ,

, 5’-

3’-

, .

, , . .

, ,

, :

. .

,

GreA

TFIIS,

, .

-

. ,

, -

, 3’-

5’-

99 . 3’-

,

.

3’. -

,

3’3’→5’-

-

E. coli.

I

.

,

,

(

)

3’-

.

,

,

, . -

,

.

,

,

GreA

GreB -

. .

-

,

, .

in vivo, , -

, (transcriptional

–

( ),

(U)

( ). 10–20

slippage),

100 . Mn2+

Mg2+

. ,

,

,

.

. ,

.

-

3

2

.

-

.

,

(A)

E. coli

(U)

10

,

dT

dA,

.

in vivo, .

,

, , ,

( )-

(

5'-

pyrBI

,

codBA). ,

, , .

, E. coli

.

101

. .

1992 . ,

.

,

-

. -

,

-

,

∼ 10 II –

(

.

.

. I.8).

I

,

.

,

:

,

. , 12-

,

–

-

∼10

-

,

.

,

,

30–40

.

, -

II I

-

I

, (

.

. I.8, ,1).

. -

3’-

-

I

. ,

-

I ( ,

-

.

. I.8, ,1). I

II

102 . ,

-

-

II I .

-

I,

. ,

,

,

. .

,

, . -

, .

, , . ,

,

, :

, , ,

(

.

. I.8, ,2),

. 3’(

.

. I.8, ,3).

3’-

,

, ,

. -

II

,

-

,

,

10

,

in vivo. . ,

II.

103 :

(general)

. , ,

,

. -

II

. I.5. P-TEFb (positive transcript elongation factor b)

SII, -

II

.

P-TEFb, I.5 -

II

,

P-TEFb

124, 43 ,

SII (TFIIS)

38 ,

TFIIF

30, 70

, (SIII) A, B 110 18 15 ELL

80

. DRB – 5,6-

-1-β-D-

DRB

104 Drosophila, – DRB.

. DRB , , , -

II .

P-TEFb

,

.

-

II,

. -

P-TEFb,

, II

. SII, ,

-

II

, . -

,

DRB-

SII

. 3’-

,

,

II

.

-

,

α-

,

–

-

,

II

.

SII

, 6-

,

,

, GTP

, UTP.

,

, -

.

,

II

105 . : (SIII)

ELL,

,

-

TFIIF,

, . ,

, .

.

, -

,

TFIIF

II 3’-OH-

,

.

,

TFIIF

3’-OH-

.

TFIIF , -

,

II

.

. (SIII) ,

A, ~150, 18

(SIII)

A-

15

B

C

. A-

,

. C,

B,

,

(SIII). (SIII)

(

-

,

) von Hippel–Lindau (VHL), .

VHL

BC,

106 .

, , BC.

ELL

(eleven–nineteen

,

lysine-rich

leukemia)

19 (19p13.1), MLL (mixed lineage

leukemia)

11 (11q23) ,

.

MLL .

"

",

, ELL,

N-

MLL.

ELL

, , 11, , . . .

,

,

. ,

(

)

. , . .

, -

,

E. coli

GC-

,

, , A,

– .

(A)-

. ,

-

GC-

107 , –

-

.

. I.9.

E. coli

,

– 3’-

-

, (U)-

(dA)-

. -

,

-

.

108 σ-

.

:

, ,

, ,

,

,

.

E. coli

,

-

ρ,

. 46

, .

,

.

,

:

ρ ,

(rut sites,

. rut – .

,

ρ-

),

.

,

-

ρ-

,

-

. E. coli ,

. ,

Trp

(

.

. I.9).

Trp

-

,

, ,

180 . .

. ,

. -

,

Trp ,

,

,

, Trp. , .

, ,

, ,

109 , .

,

2/3,

1/2

3/4

. .

ρ-

3/4

, .

(U). 3/4 . , , Trp

.

Trp

,

1, 1/2,

2/3. , . , ,

2/3, 3/4,

. . ,

–

-

I, II

III.

,

N-TEF -

II, ATP.

Reb-1

, -

I

, . Reb-1-

3’-

in vivo.

TTF-1, ,

-

110 -

I

.

L -

,

, III, . Pol I. 565 28S 21 . .

. .

. 3'-

18-

,

Sal-

(AGGTCGACCAGA/TT/ANTCCG), .

, , .

11

(GGGTCGACCAG),

. . 5'-

.

,

, , . , TTF-I

Sal-

Pol I. ,

,

,

C-

-

80

,

-

c-Myb. N,

TTF-I , .

TTF-I

Rib-1 . Pol I. , TTF-I

111 .

, Pol I, (

-

ρ-

E. coli

). Pol I,

PTRF (polymerase and transcript release factor), Pol I,

TTF-I. TTF-I

.

,

To,

170 . .

.

To

TTF-I

, . .

2.1.4. , ,

, .

, , 3’,

-

II (

.

). -

. , ,

.

MTTERM -

in vivo ,

.

MTTERM

,

-

, H-

L-

.

112

. 2.1.4. -

,

.

.

in vivo . ,

in vitro.

,

,

,

, . ,

. , . . , , .

,,

,

, , ,

.

, ,

-

,

.

:

.

113 ,

. :

,

,

1) ; 2) ,

,

; 3) . , .

,

"

"

.

.

, .

H3

H4

CAF-I (chromatin assembly factor I), 150, 60 H3

50

.

H4

, 150

,

WDTrp

(

Asn

D – ).

H2A .

ó ,

W

H2B, in vitro

H3/H4 , .

,

,

H2A/H2B, (

GAL4,

USF

Sp1).

, H1, .

in vivo

114 ,

. (

3.2.4).

,

.

Ura3 ,

Ppr1,

G2/M

.

. ,

.

,

. , , . ,

GAGA-

,

TATA. .

, . ,

, . , .

,

,

.

, 3.2.2,

,

.

,

115 , . .

.

,

-

, , . ,

TFIID,

, , ( .

3.2.2).

,

.

(HAT – histone acetyltransferase) .

Lys

N-

, (

.

. I.2).

ó .

, ,

in vitro ,

.

-

,

.

,

,

116 . . – HAT A

HAT B.

HAT A

.

.

ó HAT B ,

, . .

HAT A ,

ó HAT B

,

,

H3

H4

Lys. ,

HAT1,

-

, , HAT A. HAT A Tetrahymena

GCN5,

(Gcn5),

,

. ,

, .

Gcn5

– Ada2

Ada3, Gal4–V 16

Gen4.

Gcn5–Ada2–Ada3

-

, .

-

. Gcn5–Ada .

-

,

, . ,

-

,

117 . , , .

, ,

,

-

.

,

,

,

, ,

. Gcn5–Ada. .

HAT

(histone deacetylase – Hd), ,

.

, . - HdA (350 )

HdB (600

). HdA , .

HdB

10

HdA

,

75 HDA1

HDA3

.

Rpd3,

71

HdB ,

, ,

. HdA

Rpd3-

;

, HdA

,

,

HdB H3

H4 in vivo.

.

118 ,

,

, .

, .

:

white, .

.

,

(Lys-12

H4).

,

,

,

β. ,

, .

. –

, ,

. , . , . Swi/Snf , . NURF (nucleosome remodeling

119

I.6

Swi/Snf

NURF

,

Swi/Snf

Swi1

2000

-

Swi2 (Snf2) Swi3 Snf3 Snf5 Snf6 p78 p68 p50 p47 p25 NURF

215 140 55 38

500

,

120 factor)

(

. I.6). ,

ATP . ,

ATP

.

, . ,

Swi/Snf

,

. SWI

SNF,

,

, .

Swi/Snf, ,

, ,

.

,

. I.6, ,

(p78–p25) – .

Swi/Snf

in vitro ATP-

.

H3

H4, ,

Swi/Snf in vivo. Swi/Snf

-

II,

. ,

SWI/SNF

,

.

Swi/Snf

,

,

II

CTD-

,

Swi/Snf, . NURF GAGA, hsp70. In vivo

-

121 200–300 , TATA-

GAGA

–

HSF (heat shock factor

). , GAGAGAGA

ATP.

ATP

, NURF.

HSP70

GAGA.

NURF

,

,

,

ATP

. NURF

, ,

ATP

Swi2.

140

ATP

,

Swi2, .

,

. ATP,

, . , . ,

Swi/Snf

-

II, .

,

,

.

,

-

, . /

,

,

122

, . .

, ,

.

-

,

SP6

T7,

in vitro, ,

.

,

in vitro -

II

III,

in vivo.

,

, . ,

. -

–

,

.

. ,

.

,

. . ,

,

,

-

,

. .

123 .

,

.

,

,

.

. .

,

,

-

II,

.

, ,

,

,

. , ,

70 .

,

,

,

. , , , .

,

,

POL II

, YY1,

NMP-1,

.

,

. , ,

, .

-

, .

,

, ,

124

. , .

,

,

.

2.2.

–

.

,

,

–

-

, , .

, (

(

)

), . .

.

-

. , .

,

-

,

,

. 5’,

"

" –

5’-

. –

.

3’,

( )-

. .

,

125 , , , . . 2.2.1. ,

(

. .

. I.10, ).

. T-

3

7,

in vivo III,

.

, . III E. coli,

5S, 16S

23S .

30S

16S .

23S

,

,

.

P. , . ,

(

.

, ,

-

9).

126 . 3’. .

( )-

E. coli,

( ) 1962 .,

-

( )-

1975 .

. I.10. – I-VI; ( );

, –

,

(

– )

127 .

, , .

,

( ).

,

,

14–16

),

1

(80–200 –

40%

(∼100%

).

( )2.2.3. ( )-

. ( )( ρ-

.

. I.10, ).

3’-

,

. ,

3’-

( ).

ρ-

, , .

3’,

( ) .

( ),

–

5’-

. +

( )-

,

,

, 3’-OH-

,

-

.

, .

( )-

. ( )),

(ATP:

-

128 3’-OH: + nATP → E. coli

( )n + nPPi

( )-

.

( )-

I

pcnB, .

52

, ,

, -

, CCA-

. pcnB

E. coli.

,

pcnB

E. coli

. f310,

36,3

,

. ,

( )-

.

. 25

, .

.

,

,

,

,

.

. 4.2.2 ColE1. ,

, ColE1

pBR322,

, (

I),

129 -

(

II),

,

. I .

I

5’-

( ( )-

I–5)

–

pcnB.

I–5

, (1–2

I–5

),

( ( )-

( )--

– >10

).

I–5

.

I–5 3’-

, . ( )-

,

,

I–5

3’+

.

( )-

,

I–5 III.

, I–5, .

. . ,

pcnB,

( )-

lpp, trxA, ompA,

,

rpsO

pnp, rnb

rne, . 3’-

, ,

ρ-

(

.

. I.10). II

( )-

(

.

. I.10, ).

, ,

, ( )-

,

130

II.

, . 3’-

3’-

,

, .

,

: 3’-

"

"

.

( )+-

,

,

II

.

. I.10 , .

,

,

,

.

3’ATP-

ATP

-

,

DnaK,

, .

.

(

.

2.2.3),

( )( )-

PAB, ,

. S1, ( )-

6

3·10

–1

.

, . 30S

, ,

,

S1 . .

131 -

. .

E. coli

IV (

.

. I.10, ).

,

,

.

( )-

HeLa 55

,

– 35–55 ( )-

60

.

.

in 600 20–23

( )-

vitro

( )-

,

.

3’-

,

. ( )( ) +-

,

,

.

(75%), G (24%),

C

U(

– 5%),

, 7.

( )

( )-

(Pisum sativum)

, 43

,

( )-

,

105 . 2.2.2.

-

, (editing).

, ,

132 ,

,

,

. – Crithidia fasciculata,

:

Leishmania tarentolae (

Tripanosoma brucei.

)

20–50 ( ),

, .

(

,

), .

(U), ,

U, ,

.

"

" U(

. I.7), (

),

, (AUG),

,

. (UAG

UAA),

3’poly(A)-

.

,

.

, . , ,

-

-

,

(guide RNAs, gRNA), . g ,

5’.

15

U,

3’-

133 3’-

.

I.7

AAUUUAUGUUGUCUUU AUCUCUAAGGGUUUAACCGG  

P. polycephalum

(

AUUAAAAAGGGGCACAC 

)

CAGUAAAAAAAAAAAAAA  UUUUUCAUUGUGGUUUAC   Phe Tyr AAUAAUAUGGCGAAACAU  ,

UUUAUGCGGCAAGGA  Arg GAUAUAAUUUGAUCAGUAUA  -

5’-

g ,

,

.

,

g

. ,

,

,

. (

)

g

U. – .

,

,

-

134 – ,

, –

.

P. polycephalum (

.

. I.7). , .

.

,

.

P,

,

-

,

, (G)

,

,

.

UAA UGA,

, . .

.

,

U. : U→C. ,

,

.

,

. , ,

.

, , ,

. psbL

,

rpl2 A G

,

135 ATG.

→U.

,

, , .

,

,

,

, .

,

, ( AG),

, ,

GG. ,

,

(

1),

,

100%,

90–95%-

30-

,

.

,

, 40

80%.

,

, . , ,

.

, ,

-

. ,

,

,

,

. -

136

.

. I.8

, .

B

(APOB)

, .

APOB, APOBPOB 43

29

. . .

(

, ,

512

,

. I.11, ).

ApoB100

14

( . .).

26

UAA.

, ( ,

AA,

, )

137 I.8

,

-

/

C→U (CAAGlu→UAASTOP)

APOB-

A→I (CAGGlu→CGGArg)

» AMPA

KA

» 1

,

?

»

α-

Asp

Gly

δ » »

U→A (CUCLeu→CCCPro)

U→A (TTCPhe→TACTyr)

1

C→U (CGAArg→UGASTOP) C→U

U→C

C→U (Gly→Asp) A→I (UAGSTOP→UGGTrp) G

138

. I.11.

APOB

– APOB-

;

– ;

C –

APOBAPOBEC1. ( Z) . . APOB48 (241

),

2152 N-

APOB100. ,

-

APOB100, .

APOB-

139 .

,

,

. ,

, APOB-

,

, (

1)

28

.

,

12,

. ,

,

: , . , ,

, in vivo

in vitro.

,

,

.

55

,

25

, in vitro

25%-

. , .

,

140

Че

е

Б С К К М ш

С С С С С С С

я

1 2 1 1 1 1 1

GAUA aAaA GAUA GAUA GAUA GAUA GAUA

С→U С С С С С С С

* * ** * * * * * AAUU UGAUСAGUAUAUUAAAG AAU aUGAUСua AUuUguuua AAUU UGAUСAGUAUAUUAAAG AAUU UGAUСAGUAUAUUAAAG AAUU UGAUСAGUAUAUUAAAG AAUU UGAUСAGUAUAUUAgAG AAUU UGAUСAGUAUAUUAAAG ,

,

. , (mooring sequence). -

,

in vitro. APOB. .

,

-

, ,

APOB-

. ,

11,

, , , .

,

in vitro

C

–

3’in vivo.

,

APOB-

,

,

APOBEC1(

.

-

. I.11, ). ,

,

.

141 66

44

, ,

,

AUX240 (240

),

, . APOBEC1-

,

-

, . , ( ,

.

. I.11, ).

1 PO -

,

5’-

.

. , -

: . /

.

g

,

, ,

. -

, .

, ,

, . . ,

G,

-

,

.

,

142 APOB-

,

,

, , ,

in situ ( . .

). g

. ,

,

,

.

?

? , ?

,

,

, .

,

. , , .

5.3.1

.

,

,

. ,

,

Metazoa.

, -

,

,

.

,

,

143 -

. , ,

, . , .

,

,

-

.

" ",

,

. 2.2.3.

.

.

, -

5’-

-

, , , .

,

,

-

3’-

, 3’,

.

,

, .

-

.

144 3.3. . 5’-

,

-

.

20–30

. I.12.

.

,

–

-

,

-

-

1

. , I, II –

,

2

, .

,

eIF-4E

-

.

,

, -

,

. -

II. ,

. , , GTP,

,

– ATP, .

145 , 5’:

GMP,

ppp(A/G)pNpNpN...

5’-

5’–5’-

. : G(5’)ppp + ppp(5’)(A/G)pNpNpN... ↓ G(5’)ppp(5’)(A/G)pNpNpN... + pp + p ,

-

,

.

GTP (

–GMP.

-

GMP

γ-

5’-

),

,

.

.

-

. N7

.

. ( 7)-

-

,

S7

-

-

,

(

. I.12).

, 0-

. 2’-

5’-

(A , .

G), -

146 – 2’- -

. 1-

.

,

6

-N -

ATP,

2’- -

NH2-

.

,

2’-OH .

. I.13.

3′, .

-

( )-

, ,

, (CstF). PAP – ( ), CF I CF II –

, CPSF – I

II

-

( 1-

22.

,

.

, ,

10–15%

. I.12). ,

. 2’-

.

,

147 , (U-

,

).

2

2,2,7

m

7:

G(5')ppp(5')N.

U-

U-

-

,

,

U-

. . ,

,

3’-

, -

.

3'-

.

3’,

,

,

3’-

( )AMP: 5’ GpppG__________AAUAAA__↓________UUUUU___ 3’

↓

5’ GpppG__________AAUAAA__OH P____UUUUU___ 3’

↓

5’ GpppG__________AAUAAA__AAAAAAAAAAAAAAA 3’ : ,

.

U-

,

II. U1,

3'-

,

, , TPI (3'-terminal processing

inhibitor).

148

. I.14. PAP –

( )-

, PAB II –

, ( )-

( ) II

3’-

(

. I.13).

,

,

, .

, – AAUAAA

.

( )-

15

, U

GU.

3’-

A,

,

.

,

,

AAUAAA, , . AAUAAA

CPSF

(cleavage and polyadenylation specificity factor), .

CPSF

160, 100, 70 -

,

30

.

, .

149

AAUAAA. GUCSTF

(cleavage

stimulating

factor),

(77, 64 ).

50

GUCPSF

.

CSTF

. .

. : CFI CFII (cleavage factors).

, . , 3’-

in vitro, ( )-

–

,

. , ,

-

,

, . I.13.

β–γ-

ATP, .

, .

, .

3’. CPSF,

: CSTF, CFI (A)-

CFII. ∼ 80

∼ 43

, ,

. .

150 -

,

Ser

Thr

,

in vitro.

, .

,

. (A)-

.

( ), X,

β,

-

,

.

3’-

, AMP ,

-

-

( )-

ATP .

(A)-

CPSF,

(A)-

PAB II (poly(A)-binding protein II), ,

,

.

( )( )-

. . I.14. (

∼ 25

)

3’/

,

∼ 250

( ).

, AMP

( )-

.

, ( )-

PAB II (

. I.14).

.

PAB II

( )

( )( )

.

3’,

151 .

, . , , .

,

. I.9.

.

(1997 .) .

, ,

,

,

. , CCA

3’-

,

. ( )-

E. coli,

-

Sulfolobus shibatae, ( )-

, . , .

, ,

. I.9,

,

.

152 I.9

E. coli ( ),

80–200

14–60

∼100

2–50

,% 3’-OH-

AAUAAA : , 3’-

3’"

– "

"

–

",

S1,

PABP, ,

40S 30S

,

ColE1

153 .

, .

1977 .

,

,

. splice –

(

)

.

,

.

,

(

–

(

U6

),

.

U1–

,

.

)-

.

, , , .

(self-splicing).

, ,

,

.

, .

5’-

3’↓ ↓ ↓ 1GUAUGU__...__UACUAAC__...__(Py)nAG ←-------------------------------------------→

5’–

2–3’

, . -

.

, -

,

5’.

3’-

154 (Py)n

3'. ,

3’-

,

(branch point). 5’-

, "

" (lariat) (

.

).

,

-

, .

I, II ,

–

III,

,

.

I, II

III.

,

-

–

,

(

.

9.2). , .

,

: −

−

  R–OH + R’–O–P–OR’’ → R–O–P–OR’’ + R’–OH,

I, II . I.15, , .

III

. I–III

I .

Tetrahymena

thermophila

. I.10

.

155 I.10

I

GTPOH + [

1]upA−

GTPOH −Gpa[

+ 2]

[

1]upA−

−Gpa[

2]

5’1]u−OH −Gpa[

[ GpA−

1]u−OH −Gpa[

[ 2]

GpA−

2]

2 [

1]upa[ + GpA−

2] −G

[

1]upa[ + GpA−

2] −G

156

157

. I.15. – thermophila), II –

I-III I (Tetrahymena III ,

(Euglena gracilis). ,

3′, , EBS ; – II; –

IBS –

5′-

158 . , 1, 2 – , ,

–

.

.

I

,

II

(maturase,

. mature –

), in vivo. I OH-

2’-

GTP

5’-

. I.10) (

(

.

GTP ).

5’-

.

1.

3’-OH-

1

3’-

, . .

"

.

" 3’-OH-

, 2.

3’-OH-

1 ,

G,

1

. II (

.

III

. I.15, ).

.

2’-OH-

,

, ,

5’-

,

1 .

5’-

2’–5’,

3’-

OH-

. 1

3’-

.

159

5’-

. . , . -

. , ,

II

III

,

. ,

,

.

. .

–

ATP

,

, .

,

, , .

OH-

5’.

II

III

,

. -

.

(

)

: 3’-

5’-

. ,

,

, . -

5’U1-

65

U2AF ,

U1, 5’-

, 35

U2AF ,

160 3’-

.

. , , Ser

SR-

Arg,

70 -

U1.

-

(70

),

U2AF35

,

,

SR-

"

,

,

". 5’-

,

3’-

. 5’U2AF

U1-

,

∼ 137

,

,

3’,

"

".

. .

, (

. .),

, 100 . . .

,

,

, , .

(

,

)

,

,

.

, , . ,

.

-

,

,

161

. I.16. –

; (2),

–

–

(1) -

(CBC)

. :1–

,2

, 3–

,X–

,

162 ;+– ; AAUAAA –

, ( )-

,

,

.

,

-

,

, ,

( U1, U2AF

. I.16, ).

U2-

,

SC35,

,

3’-

–

5’-

–

ASF/SF2. , .

,

,

. ,

, (

.

-

,

. I.16, ). .

,

,

,

:

,

,

;

(

)

;

, -

,

. 51, 32, 11

6%.

,

, .

. ,

,

1600

3,5%

1% – 400 50

,

.

300 ,

,

, in vitro

. ,

163 400

50

. .

,

6

. .,

7

, , . .

2.2.4.

-

, in vivo

II .

– – (

).

-

,

. , ,

, ,

.

, ,

,

, – ( )-

PABP

3.4.

50.

,

,

-

. -

(CBC).

-

, (

)

, ,

.

-

CBC

164 (cap-binding complex).

CBC

CBP80 (

80

)

CBP20 (20

). N-

CBC NLS (nuclear localization signal), CBC

( CBC

.

).

-

, -

.

CBC –

. CBC

-

.

15

,

m7G

-

.

,

-

.

, CBC – CBP80. ,

CBC -

.

,

-

.

. I.16

, . -

.

(

,

5'-

.

. I.16, ,2).

-

3',

,

(

I.16, ,3).

-

.

,

,

CBC.

,

3'-,

5'-

, CBC (

.

) (

165 ).

5'-

CBC-

(

(

.

. I.16, ,4). ,

,

.)

, ,

,

5'-

-

.

, U1-

CBC

5'-

,

, ,

, . 3'-

-

.

,

,

3'.

, -

( )-

7

-

m GpppG

CBC

,

.

,

-

CBC

,

3'-

,

–

,

. 5'-

3'(35–40 . .)

. ,

5',

( )-

-

.

,

,

3'-

,

, .

CBC .

-

II

II ,

,

166 : -

U-

.

,

.

-

.

in situ

,

,

CBC

, Chironomus tentans.

(35–40 . . .) , ,

,

200 ,

.

CBC .

,

, .

,

CBC CBP80 –

(NLS). α (

– 60

β

)

(90 NLS

,

α

).

,

,

,

, NPC (nuclear pore complex). .

NLS,

NPC

,

, . , GTP

Ran,

GTP, α.

, . , NPC. α

NLS

. α

CBC, .

, NLS

, α

167 -

CBC,

α,

NPC

. CBC–

NPC α

–

α.

GTP

,

Ran

GDP

1,

GTP

-

Ran (Ran GAP1), GTP.

NLS

-

CBC, CBC-

.

, .

-

U–

, .

eIF-

4E

( CBC

.

2.5.1).

.

.

. -

( . . )

5'-

CBC

.

-

,

, ,

,

-

.

-

I,

-

I

II

III.

,

III,

5'-

, m7G-

, . .

,

,

168 ,

,

,

. , . 2.3.

, .

,

,

, .

-

,

,

.

,

-

,

, .

("

")

, ,

,

.

, . ,

.

, ,

, .

,

. (

169 )

. .

2.3.1. 1.3

, .

,

. ,

,

3.2.4.

,

,

.

, -

, ,

. , .

,

.

, .

, .

-

, ,

.

,

170 . ( .

, 1885 .)

, . , Schizosaccharomyces pombe

G2

. , S. cerevisiae

,

,

– Sir3

Sir4

(silent information regulators), , ,

, .

. ,

.

,

. .

(1993 .), .

,

,

–

neu

50% –

,

.

,

,

, . ,

,

,

,

. ,

, (

.

.

).

(1993 .),

, ,

,

, .

,

171 , . ,

-

. 2.3.2. -

(nucleolus)

. , . , ,

.

, , –

, .

,

250 44

. . .

, .

. : (1), (3)

(2)

. , ,

-

I,

UBF

I.

, , .

,

,

172 ,

. 28S

18S

,

, .

15–20 ,

-

.

,

, . -

,

. ,

"

" , , -

I .

,

.

,

. .

,

. ,

. ~200

.

: , ~20% . ,

,

, , . ,

.

173 . "

" (cellular aging).

,

, ,

Cdc 14,

.

, RENT (regulator of nucleolar silencing and telophase exit), ,

,

, .

2.3.3.

, . , , , ,

,

. . ,

-

,

, . 3–20 -

.

, .

, .

" "

"

" , .

174

, .

,

,

,

,

,

.

,

,

.

, -

. . , . , 20–50

,

.

, .

,

, 4%

.

,

, .

,

,

-

. , (NPC).

NPC

, ,

.

,

, ( -

,

), .

175 ,

-

,

. .

, ,

. Chironomus tentans, .

,

.

-

,

,

,

,

,

, .

, ,

5’-

.

,

.

-

,

.

, .

, , . (gene gating model),

.

(1985 .), (

)

, . , , .

.

176 , ,

,

– .

, ,

,

.

, in vitro

∼1

/

. in vivo,

50

/

.

,

, . 2.3.4. ,

. ,

.

(coiled bodies).

,

,

. , , U3.

, (

U7-

,

p80). .

: G1

. .

177 . U73’-

-

,

.

,

, -

.

PML.

(PML) t(15;17) (

15

17),

α-

PML

. 2+

PML,

Zn -

"

,

RING",

.

U1-

80 ,

,

.

PML . , , . PML-

. , ,

PML.

, . ,

PML

. WT1.

WT1

-

, ,

.

,

, , ,

,

. -

178 "

" (

Pro/Gln.

.

2.2.2)

N-

,

WT1

GC, .

.

WT1

,

, . 2.3.5.

.

,

-

,

,

(

-

,

), ,

-

,

. ,

,

,

. , .

(

.

4). , .

, ,

, . ,

. ,

,

,

,

,

. , , ;

; .

:

2)

3) ,

,

1)

179 -

II. ,

(

)

(

)

, .

(

),

4. .

.

,

,

, ,

.

,

. . .

( .

,

, 1996 .), .

,

, .

, ,

(

:

.

5.3).

, , , . . .

180 2.4. ,

, .

,

, ,

. .

, ,

,

. .

, (

),

.

, .

,

, .

, . . , ,

,

, .

.

. 2.4.1. ∼ 2,5

,

,

181 . . 80S

, 70S. :

50S,

– 40S

– 30S

60S. 70S

55–60

,

80S

75–85. , . E. coli.

50 .

5

,

– 61

, 10–20

E. coli.

.

21 350

.

30S

16S

1542

(

). 850

.

, ∼ 1450

34 ,

–

23S (2904

)

5S

(120

).

,

,

,

.

. 23S CCA-

,

. 30S

50S

,

,

. .

182 , :

-

,

–

.

,

,

.

. I.17.

50S

E. coli

in vitro , ( ,

) .

. , . 70S . 50S T. thermophilus

3 Å, 7–20 Å.

,

183 ,

10

, , .

"

": ,

.

,

–

.

184

. I.18. E. coli ( ) 30S-

, ( ) ,

"

" (stalk) 50S ,

-, -

-

,

L1

. –

16S

185 ,

–

,

,

–

(

).

. ,

.

,

.

, , . ,

,

,

.

, .

, .

,

.

. ,

2-

,

, 5–10 Å

.

186 ,

,

.

, 2+

2+

(Mg

Mg2+

Ca ).

,

.

,

. . I.17

E. coli , ,

.

. ,

. . .

, ,

.

in vitro

. ,

,

. , (2H

, 3

). , , , .

,

187 ,

. 93

. ,

, .

. , .

,

, 16S

,

. .

.

, . . I.18 70S

E. coli,

.

, . 2.4.2.

1960-

., . E. coli, . ,

(

)

188 ,

:

,

. 70S

80S N.

,

.

, . ,

.

5’3’-

,

.

N-

.

E. coli. . 30S

,

,

, (

, AUG)

.

30S – IF1, IF2

IF3.

GTP, (

.

IF2

. I.19).

,

30S

GTP, ( ,

(fMet)-

fMet

E. coli,

). .

Met

MetE. coli

30S

fMet

,

NH2,

Met.

. I.19 (

, ’, ,

189 ’).

. I.19.

E. coli , GTP, fMet-

fMet

30S AUG-

(

’

B’). (

.

C),

C (

)

, . IF1

IF3

50S

, (

D).

GTP

GDP

IF2 ( -

fMet

E).

AUG(P)

,

(A) .

190

– . .

, ,

70S

,

, ,

, .

C-

, .

. I.20. ,

191 EF-Tu,

GTP (

.

A-

. I.20,

1).

A. , A-

,

.

(

GTP

EF-Tu•GDP

2) NH2, -

A-

(

.

3).

GDP

GTP

Tu•GDP

EF-

EF-Ts.

,

A-

,

E-

(

. exit –

). -

.

A-

.

P-

( 4)

. EF-G, GTP.

,

GTP

. EF-G

.

A-

. A( fMet

-

∼ 20 .

.

. P-

E-

, .

200

∼10

A-

1),

, ,

192 P-

E-

.

A

E

,

,

A-

E-

,

. EF-G.

,

IV EF-G EF-Tu. EF-G ,

ó

EF-Tu ,

-

.

-

EF-G

. IV

,

EF-G A-

,

.

, .

. , (

A-

)

P-

,

-

.

RF1

RF2 .

RF3,

EF-G,

EF-

Tu,

-

. ,

.

, .

, E. coli

-

. – ,

–

193 .

,

EF-G

(RF)

GTP , .

RF4 (

RRF –

ribosome-recycling factor)

P-

.

E-

(

,

)

-

,

RF-

A-

.

.

IF3.

,

, -

,

. . ,

, . E. coli .

,

, , . ( ,

) .

,

E. coli,

,

. (

,

,

AUGA). ,

SD-

, (

,

194 ). .

, , .

,

. E. coli

UUG

, . IF3

, . ,

, (RF).

R17, ,

-

7

,

RF4 , , .

N,

,

-

7

-

RF4

,

. .

.

, (hybrid states model)

A-

P-

30S A-, P-

. -

E-

–

EF-

Tu•GTP• A/E.

A-

195 30S

,

CCA-

,

EF-Tu,

,

E-

,

.

GTP

EF-Tu,

CCA, -

-

A-

A/A-

,

A-

.

,

P-

,

E-

. A/P-

,

:

A-

CCA-

30S

,

P-

. P/E-

P-

:

,

CCA-

.

EF-G

EGTP,

,

30S

.

-

P/P-

,

P-

,

EE-

.

, .

-

, ,

.

-

,

:

, 30S

,

,

,

.

-

,

.

, .

, ,

196

, S21

L11.

, , . (displacement model). , , . .

–

. , ,

,

. ,

,

EF-Tu-

GTP .

GTP (cognate)

(noncognate)

∼ 104

,

.

EF-Tu•GDP -

-

-

,

.

, GTP

. ,

, .

, ,

A-

"

A-

"

vice versa. PA-

:

ram-

.

,

197 P-

,

.

"

"

,

, A-

P-

, . 10Sa

,

.

,

3’-

, .

10Sa

,

,

,

,

E. coli

ssrA, .

Ala

3’-

,

,

, . 10Sa

Ala

,

,

G3:U70, Ala

:

.

Ala

10Sa

.

,

, – ANDENYALAA –

,

CUAA-

10Sa

, , . 2.4.3.

,

. I.21 ,

. ,

.

,

. C-

198 ,

. ∼7500

,

∼4500

. . ,

(

) . .

. I.21.

–

,

199 . 3’. , -

A-

. ,

-

.

C,

. ,

. . 50S

. -

,

.

, .

, -

,

. A-

50S

, -

A-

,

,

. .

.

-

P-

. . –

, .

, ,

EF2(EF-G), GTP-

, .

EF2(EF-G)

GTP

GTP.

-

,

200 . . . 30S

,

-

30S

, .

,

30S

. . (

) 30S

,

. -

,

, A-

. . -

. .

,

. 2.5. ,

.

, ,

, , . ,

.

( .

7.4).

201

,

.

,

.

, ,

,

S. cerevisiae. 2.5.1.

, , , ,

. ,

,

,

3'-

( ),

,

-

.

,

, ,

. , .

,

(

), ,

5'-

(5'UTR – 5' untranslated

region),

.

, ( )-

,

3'-

(3'UTR).

.

5'UTR,

,

"

"

-

(uORF – upstream open reading frame), (

).

5'UTR , .

5'UTR

.

202 (

) .

3'UTR

( ). , 3'-

,

( )-

. 2.5.2. , ,

,

. (

)

5'AUG-

,

5'UTR. AUG-

.

,

,

5'-

, ,

, . .

, ,

.

.

-

,

, 5'-

3'-

, 11

25

(

,

. I.11). I.11 S. cerevisiae

eIF1

Met-

AUG-

(

)

40S

203 eIF1A

40S–60S ,

Met(

eIF2

α, ,

eIF2B

α, , , ,

eIF3

α, , , ,

)

eIF2 Met-

, , ,

40S

40S–60S

eIF4A

,

eIF4B

,

eIF4E

-

eIF4G

G1(p150),

eIF3, eIF4E

Pab 1p

G2(p130) eIF4H

eIF4B

eIF4F

eIF5 eIF5A

,

eIF6

40S–60S

, , , ,

. . 5'-

-

-

(CBC).

CBC

.

,

,

2.2.4,

CBC , .

204 eIF2

Met-

.

eIF2 Met-

vitro.

in .

GTP

eIF2

MetMet-

–eIF2–GTP.

eIF2B

GDP

, GTP

eIF2–GDP. . ,

40S 40S-

.

, –

,

. -

eIF4E

.

-

.

eIF4E

eIF4G

eIF4F. ,

eIF4E -

eIF4F,

eIF4G

. ,

eIF4A,

eIF4G

.

eIF4F

ATP,

eIF4B.

eIF4H

eIF4F

(1998

.)

eIF4B, .

eIF4F: eIF4G

eIF4E,

eIF4G1.

p20,

( )-

Pab 1p,

p20

BP –

4E-

. eIF4G eIF3, eIF4E

eIF4A.

, -

,

eIF4G

205 eIF4F.

eIF4E (

)

3.4.1. , –

–

, -

,

eIF4G

eIF3.

,

, ,

.

40S

MetMet

–eIF2–GTP,

. ,

40S

,

,

GTP

Met-

eIF3

–eIF2–

eIF4G.

,

,

eIF4F,

eIF4G

eIF4E,

-

-

.

,

eIF4A,

,

eIF1A. 5'UTR AUG-

.

3'-

( )-

.

– , N-

eIF4G

,

( )3'-

Pab1p.

( )60S

,

pab1. , 3'-

( ).

, .

,

206 . , AUG-

,

-

,

.

5'UTR

(

,

SD-

),

TIR

(translation initiation region),

3'-

16S

. .

, 5'UTR . . , -

5'UTR, " AUG-

"

.

,

.

-

SDAUG–

Met-

,

.

.

, ATP

-

5'UTR.

, eIF4A,

, ,

-

eIF4B.

,

, 5'UTR, .

,

207 AUG-

eIF2.

, . ,

eIF5 .

,

AUG IF3. eIF5,

in vivo

UUG. eIF2

,

eIF5 ß-

eIF2

eIF5. 40S 60S eIF5, . , GTP.

,

eIF2–GDP, ,

eIF1A

eIF3.

, . . . AUG,

AUG-

.

,

. A –3

AUG (

A

AUG-

+1),

. ,

,

A-

AUG,

,

, GC. 5'UTR

,

208 GC-

A,

,

.

,

AUG

,

-

.

,

: AACAATGGC. –3

G

+4. +5 ,

+6.

,

.

,

, (leaky scanning).

3.4.1 (

.

. I.40, – ) . ,

AUG-

,

5'UTR.

,

5'-

AUG

.

5' 3'.

5'UTR

AUG,

,

(uAUG, uORF) . . ,

uAUG

5'-

15–20

.

AUG. , ,

,

, .

. ,

AUG.

209 , AUG. CUG – AUC

ACG.

-AUG.

eIF2 β

∼ 10%

γ.

AUU

.

AUG,

.

,

,

.

,

,

.

,

. ,

N-

,

, . . 5'UTR

40S

. 5'-

.

,

– /

–30 ,

5'-

52

.

5'UTR,

, -

, 85%,

5'-

.

, –50

/

.

,

210 ,

– AUG-

.

,

, , ,

.

,

,

14

, AUG,

,

AUG. .

,

35S

(CaMV),

, , (

.

. I.40, ).

, . , , . , .

,

,

,

in trans,

. .

, .

,

,

.

, ,

. ,

, . .

,

,

,

, . .

.

, .

,

5'UTR

211 ,

, .

. ,

, .

,

3.4.

, : , . ,

,

. ,

,

, . AUG-

.

IRES-

(internal ribosome entry

site). AUG-

, TIR, IRES,

. .

.

, AUG-

,

-

.

, eIFG eIF4E – .

N-

, -

CeIF3

eIF4A.

212 (

)

-

-

,

IRES-

.

2.5.3.

,

, .

, ,

. . eEF1A, EF1A(EF-Tu). ,

GTP

-

-

.

eEF1B

eEF2 EF1B(EF-Ts)

EF2(EF-G)

.

eEF1B,

, eEF1A–GDP.

,

GDP

GTP

eEF2,

, -

E-

P-

. ( eEF1A

.

3.4).

eEF2 .

,

,

. , ,

,

, , .

213 ,

.

, , ,

.

,

,

-

,

.

E. coli, .

.

(

.

3.6.1), , . eEF-3

. eEF3,

. 116

eEF1A•

,

-

ATP

•GTP

.

-

,

-

. ATP,

. , ,

, . ,

eEF3

. 2.5.4.

,

,

-

214 .

.

, RRF/RF4.

.

, -

,

, -

(RF),

eRF1

eRF3.

eRF1 (UAA, UAG

UGA)

eRF3

.

GTP

,

eEF1A,

,

GTP, . .

. C. : UAA(53%) > UGA(27%) > UAG(20%). ,

UAA

– 87%. ,

. lacZ

: G>U>A>C (UGA), G>A>U>C (UAA)

A>U>C>G (UAG).

. eRF-

. , , , .

215 E. coli:

, . UGA

UUC

UCC(Ser),

UUU(Phe),

UUA –

AAG(Lys).

P-

,

RF-

,

-

A-

.

E. coli

. 2.5.5. , , , ATP.

– (

),

. , α-

(

Rickettsia prowazekii), -

-

.

,

,

( 13

)

, 22

.

. ( 16569

. .), .

-

,

-

-

,

.

.

216 ,

.

–

–

,

.

,

.

,

N-

, . .

, . . TGA

Trp, .

AGR (

R=A

,

G)

,

Ser,

,

, – Arg.

, .

, 32

, . ,

22

13

. U

(wobble) , .

. , ,

, .

,

.

217 ,

,

. ,