Alternative transcription cycle for bacterial RNA polymerase

Timothy T. Harden, Karina S. Herlambang, Mathew Chamberlain, Jean-Benoît Lalanne, Christopher D. Wells, Gene-Wei Li, Robert Landick, Ann Hochschild, Jane Kondev () and Jeff Gelles ()
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Timothy T. Harden: Brandeis University
Karina S. Herlambang: Brandeis University
Mathew Chamberlain: Brandeis University
Jean-Benoît Lalanne: Massachusetts Institute of Technology
Christopher D. Wells: Blavatnick Institute, Harvard Medical School
Gene-Wei Li: Massachusetts Institute of Technology
Robert Landick: University of Wisconsin
Ann Hochschild: Blavatnick Institute, Harvard Medical School
Jane Kondev: Brandeis University
Jeff Gelles: Brandeis University

Nature Communications, 2020, vol. 11, issue 1, 1-11

Abstract: Abstract RNA polymerases (RNAPs) transcribe genes through a cycle of recruitment to promoter DNA, initiation, elongation, and termination. After termination, RNAP is thought to initiate the next round of transcription by detaching from DNA and rebinding a new promoter. Here we use single-molecule fluorescence microscopy to observe individual RNAP molecules after transcript release at a terminator. Following termination, RNAP almost always remains bound to DNA and sometimes exhibits one-dimensional sliding over thousands of basepairs. Unexpectedly, the DNA-bound RNAP often restarts transcription, usually in reverse direction, thus producing an antisense transcript. Furthermore, we report evidence of this secondary initiation in live cells, using genome-wide RNA sequencing. These findings reveal an alternative transcription cycle that allows RNAP to reinitiate without dissociating from DNA, which is likely to have important implications for gene regulation.

Date: 2020
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DOI: 10.1038/s41467-019-14208-9

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