Molecular mechanism of transcription inhibition by peptide antibiotic Microcin J25

Karen Adelman, Julia Yuzenkova, Arthur La Porta, Nikolay Zenkin, Jookyung Lee, John T. Lis, Sergei Borukhov, Michelle D. Wang, Konstantin Severinov

Research output: Contribution to journalArticlepeer-review

121 Scopus citations

Abstract

21 amino acid peptide Microcin J25 (MccJ25) inhibits transcription by bacterial RNA polymerase (RNAP). MccJ25-resistance mutations cluster in the RNAP secondary channel through which incoming NTP substrates are thought to reach the catalytic center and the 3′ end of the nascent RNA is likely to thread in backtracked transcription complexes. The secondary channel also accepts transcript cleavage factors GreA and GreB. Here, we demonstrate that MccJ25 inhibits GreA/GreB-dependent transcript cleavage, impedes formation of backtracked complexes, and can be crosslinked to the 3′-end of the nascent RNA in elongation complexes. These results place the MccJ25 binding site within the secondary channel. Moreover, single-molecule assays reveal that MccJ25 binding to a transcribing RNAP temporarily stops transcript elongation but has no effect on the elongation velocity between pauses. Kinetic analysis of single-molecule data allows us to put forward a model of transcription inhibition by MccJ25 that envisions the complete occlusion of the secondary channel by bound inhibitor.

Original languageEnglish (US)
Pages (from-to)753-762
Number of pages10
JournalMolecular Cell
Volume14
Issue number6
DOIs
StatePublished - Jun 18 2004
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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