Structural basis for transcription inhibition by E. coli SspA.

Abstract:

:Stringent starvation protein A (SspA) is an RNA polymerase (RNAP)-associated protein involved in nucleotide metabolism, acid tolerance and virulence of bacteria. Despite extensive biochemical and genetic analyses, the precise regulatory role of SspA in transcription is still unknown, in part, because of a lack of structural information for bacterial RNAP in complex with SspA. Here, we report a 3.68 Å cryo-EM structure of an Escherichia coli RNAP-promoter open complex (RPo) with SspA. Unexpectedly, the structure reveals that SspA binds to the E. coli σ70-RNAP holoenzyme as a homodimer, interacting with σ70 region 4 and the zinc binding domain of EcoRNAP β' subunit simultaneously. Results from fluorescent polarization assays indicate the specific interactions between SspA and σ70 region 4 confer its σ selectivity, thereby avoiding its interactions with σs or other alternative σ factors. In addition, results from in vitro transcription assays verify that SspA inhibits transcription probably through suppressing promoter escape. Together, the results here provide a foundation for understanding the unique physiological function of SspA in transcription regulation in bacteria.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Wang F,Shi J,He D,Tong B,Zhang C,Wen A,Zhang Y,Feng Y,Lin W

doi

10.1093/nar/gkaa672

subject

Has Abstract

pub_date

2020-09-25 00:00:00

pages

9931-9942

issue

17

eissn

0305-1048

issn

1362-4962

pii

5891570

journal_volume

48

pub_type

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