E. coli TraR allosterically regulates transcription initiation by altering RNA polymerase conformation

被引：54

作者：

Chen, James ^{[1
]}

Gopalkrishnan, Saumya ^{[2
]}

Chiu, Courtney ^{[1
]}

Chen, Albert Y. ^{[2
]}

Campbell, Elizabeth A. ^{[1
]}

Gourse, Richard L. ^{[2
]}

Ross, Wilma ^{[2
]}

Darst, Seth A. ^{[1
]}

机构：

[1] Rockefeller Univ, 1230 York Ave, New York, NY 10021 USA

[2] Univ Wisconsin, Dept Bacteriol, Madison, WI 53706 USA

来源：

ELIFE | 2019年 / 8卷

基金：

美国国家卫生研究院;

关键词：

OPEN COMPLEX-FORMATION; ESCHERICHIA-COLI; RIBOSOMAL-RNA; CRYO-EM; CRYSTAL-STRUCTURE; STRUCTURAL BASIS; ALPHA-SUBUNIT; TERMINAL DOMAIN; PPGPP BINDING; PROMOTER;

D O I：

10.7554/eLife.49375

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

TraR and its homolog DksA are bacterial proteins that regulate transcription initiation by binding directly to RNA polymerase (RNAP) rather than to promoter DNA. Effects of TraR mimic the combined effects of DksA and its cofactor ppGpp, but the structural basis for regulation by these factors remains unclear. Here, we use cryo-electron microscopy to determine structures of Escherichia coli RNAP, with or without TraR, and of an RNAP-promoter complex. TraR binding induced RNAP conformational changes not seen in previous crystallographic analyses, and a quantitative analysis revealed TraR-induced changes in RNAP conformational heterogeneity. These changes involve mobile regions of RNAP affecting promoter DNA interactions, including the Globe, the clamp, the bridge helix, and several lineage-specific insertions. Using mutational approaches, we show that these structural changes, as well as effects on sigma(70) region 1.1, are critical for transcription activation or inhibition, depending on the kinetic features of regulated promoters.

引用

页数：29

共 87 条

[11] Structures of an RNA polymerase promoter melting intermediate elucidate DNA unwinding [J].