Binding interface between the Salmonella σ(S)/RpoS subunit of RNA polymerase and Crl: hints from bacterial species lacking crl

Scientific Reports
Paola CavaliereF Norel

Abstract

In many Gram-negative bacteria, including Salmonella enterica serovar Typhimurium (S. Typhimurium), the sigma factor RpoS/σ(S) accumulates during stationary phase of growth, and associates with the core RNA polymerase enzyme (E) to promote transcription initiation of genes involved in general stress resistance and starvation survival. Whereas σ factors are usually inactivated upon interaction with anti-σ proteins, σ(S) binding to the Crl protein increases σ(S) activity by favouring its association to E. Taking advantage of evolution of the σ(S) sequence in bacterial species that do not contain a crl gene, like Pseudomonas aeruginosa, we identified and assigned a critical arginine residue in σ(S) to the S. Typhimurium σ(S)-Crl binding interface. We solved the solution structure of S. Typhimurium Crl by NMR and used it for NMR binding assays with σ(S) and to generate in silico models of the σ(S)-Crl complex constrained by mutational analysis. The σ(S)-Crl models suggest that the identified arginine in σ(S) interacts with an aspartate of Crl that is required for σ(S) binding and is located inside a cavity enclosed by flexible loops, which also contribute to the interface. This study provides the basis for further structural invest...Continue Reading

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Citations

Dec 13, 2017·The Biochemical Journal·Paola CavaliereFrançoise Norel
Sep 6, 2019·Proceedings of the National Academy of Sciences of the United States of America·Alexis Jaramillo CartagenaSeth A Darst
Oct 13, 2020·Frontiers in Microbiology·Herb E Schellhorn

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Methods Mentioned

BETA
X-ray
two hybrid
isothermal titration calorimetry
NMR
chemical shift

Software Mentioned

CYANA
CCPNMR
ZDock
RosettaDock
Haddock Webserver
PyMOL
ElNémo
NMRPipe
Topspin

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