Rxivist logo

Chemical-genetic interrogation of RNA polymerase mutants reveals structure-function relationships and physiological tradeoffs

By Anthony Lyndon Shiver, Hendrik Osadnik, Jason M. Peters, Rachel A. Mooney, Peter I Wu, James C Hu, Robert Landick, Kerwyn Casey Huang, Carol A. Gross

Posted 17 Jun 2020
bioRxiv DOI: 10.1101/2020.06.16.155770

The multi-subunit bacterial RNA polymerase (RNAP) and its associated regulators carry out transcription and integrate myriad regulatory signals. Numerous studies have interrogated the inner workings of RNAP, and mutations in genes encoding RNAP drive adaptation of Escherichia coli to many health- and industry-relevant environments, yet a paucity of systematic analyses has hampered our understanding of the fitness benefits and trade-offs from altering RNAP function. Here, we conduct a chemical-genetic analysis of a library of RNAP mutants. We discover phenotypes for non-essential insertions, show that clustering mutant phenotypes increases their predictive power for drawing functional inferences, and illuminate a connection between transcription and cell division. Our findings demonstrate that RNAP chemical-genetic interactions provide a general platform for interrogating structure-function relationships in vivo and for identifying physiological trade-offs of mutations, including those relevant for disease and biotechnology. This strategy should have broad utility for illuminating the role of other important protein complexes. ### Competing Interest Statement The authors have declared no competing interest.

Download data

  • Downloaded 366 times
  • Download rankings, all-time:
    • Site-wide: 91,540
    • In systems biology: 2,049
  • Year to date:
    • Site-wide: 74,371
  • Since beginning of last month:
    • Site-wide: 55,644

Altmetric data

Downloads over time

Distribution of downloads per paper, site-wide