Structure and function of the transcription elongation factor GreB bound to bacterial RNA polymerase

被引:167
作者
Opalka, N
Chlenov, M
Chacon, P
Rice, WJ
Wriggers, W
Darst, SA
机构
[1] Rockefeller Univ, New York, NY 10021 USA
[2] Scripps Res Inst, Dept Biol Mol, La Jolla, CA 92037 USA
[3] NYU, Med Ctr, Skirball Inst Biomol Med, New York, NY 10016 USA
[4] NYU, Med Ctr, Dept Cell Biol, New York, NY 10016 USA
基金
美国国家卫生研究院;
关键词
D O I
10.1016/S0092-8674(03)00600-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Bacterial GreA and GreB promote transcription elongation by stimulating an endogenous, endonucleolytic transcript cleavage activity of the RNA polymerase. The structure of Escherichia coli core RNA polymerase bound to GreB was determined by cryo-electron microscopy and image processing of helical crystals to a nominal resolution of 15 Angstrom, allowing fitting of high-resolution RNA polymerase and GreB structures. In the resulting model, the GreB N-terminal coiled-coil domain extends 45 Angstrom through a channel directly to the RNA polymerase active site. The model leads to detailed insights into the mechanism of Gre factor activity that explains a wide range of experimental observations and points to a key role for conserved acidic residues at the tip of the Gre factor coiled coil in modifying the RNA polymerase active site to catalyze the cleavage reaction. Mutational studies confirm that these positions are critical for Gre factor function.
引用
收藏
页码:335 / 345
页数:11
相关论文
共 81 条
[71]   SPONTANEOUS CLEAVAGE OF RNA IN TERNARY COMPLEXES OF ESCHERICHIA-COLI RNA-POLYMERASE AND ITS SIGNIFICANCE FOR THE MECHANISM OF TRANSCRIPTION [J].
SURRATT, CK ;
MILAN, SC ;
CHAMBERLIN, MJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (18) :7983-7987
[72]   Crystal structure of a yeast TFIIA/TBP/DNA complex [J].
Tan, S ;
Hunziker, Y ;
Sargent, DF ;
Richmond, TJ .
NATURE, 1996, 381 (6578) :127-134
[73]   GreA and GreB proteins revive backtracked RNA polymerase in vivo by promoting transcript trimming [J].
Toulmé, F ;
Mosrin-Hauman, C ;
Sparkowski, J ;
Das, A ;
Leng, M ;
Rahmouni, AR .
EMBO JOURNAL, 2000, 19 (24) :6853-6859
[74]   Basic mechanisms of transcript elongation and its regulation [J].
Uptain, SM ;
Kane, CM ;
Chamberlin, MJ .
ANNUAL REVIEW OF BIOCHEMISTRY, 1997, 66 :117-172
[75]   Crystal structure of a bacterial RNA polymerase holoenzyme at 2.6 Å resolution [J].
Vassylyev, DG ;
Sekine, S ;
Laptenko, O ;
Lee, J ;
Vassylyeva, MN ;
Borukhov, S ;
Yokoyama, S .
NATURE, 2002, 417 (6890) :712-719
[76]   IDENTIFICATION OF A 3' -] 5' EXONUCLEASE ACTIVITY ASSOCIATED WITH HUMAN RNA POLYMERASE-II [J].
WANG, DG ;
HAWLEY, DK .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (03) :843-847
[77]   Modeling tricks and fitting techniques for multiresolution structures [J].
Wriggers, W ;
Chacón, P .
STRUCTURE, 2001, 9 (09) :779-788
[78]   In vitro characterization of mutant yeast RNA polymerase II with reduced binding for elongation factor TFIIS [J].
Wu, JS ;
Awrey, DE ;
Edwards, AM ;
Archambault, J ;
Friesen, JD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (21) :11552-11557
[79]   MECHANISTIC STUDIES ON DEOXYRIBONUCLEIC-ACID DEPENDENT RIBONUCLEIC-ACID POLYMERASE FROM ESCHERICHIA-COLI USING PHOSPHOROTHIOATE ANALOGS .1. INITIATION AND PYROPHOSPHATE EXCHANGE-REACTIONS [J].
YEE, D ;
ARMSTRONG, VW ;
ECKSTEIN, F .
BIOCHEMISTRY, 1979, 18 (19) :4116-4120
[80]   Crystal structure of Thermus aquaticus core RNA polymerase at 3.3 Å resolution [J].
Zhang, GY ;
Campbell, EA ;
Minakhin, L ;
Richter, C ;
Severinov, K ;
Darst, SA .
CELL, 1999, 98 (06) :811-824