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

Chen, J.; Gopalkrishnan, S.; Chiu, C.; Chen, A. Y.; Campbell, E. A.; Gourse, R. L.; Ross, W.; Darst, S. A.

doi:10.1101/766725

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bioRxiv · 10.1101/766725

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

Chen, J.·Gopalkrishnan, S.·Chiu, C.·Chen, A. Y.·Campbell, E. A.·Gourse, R. L.·Ross, W.·Darst, S. A.

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Abstract

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. How TraR and its homologs regulate transcription is 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 of RNAP conformational heterogeneity revealed TraR-induced changes in RNAP dynamics. These changes involve mobile regions of RNAP affecting promoter DNA interactions, including the {beta}lobe, 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.

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Chen, J., Gopalkrishnan, S., Chiu, C., Chen, A. Y., Campbell, E. A., Gourse, R. L., Ross, W., Darst, S. A.. 2019-09-12. E. coli TraR allosterically regulates transcription initiation by altering RNA polymerase conformation and dynamics. https://doi.org/10.1101/766725

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