Temporal regulation of gene expression of the Escherichia coli bacteriophage phiEco32

Olga Pavlova, Daria Lavysh, Evgeny Klimuk, Marko Djordjevic, Dmitry A. Ravcheev, Mikhail S. Gelfand, Konstantin Severinov, Natalja Akulenko

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Escherichia coli phage phiEco32 encodes two proteins that bind to host RNA polymerase (RNAP): gp79, a novel protein, and gp36, a distant homolog of σ70 family proteins. Here, we investigated the temporal pattern of phiEco32 and host gene expression during infection. Host transcription shutoff and three distinct bacteriophage temporal gene classes (early, middle, and late) were revealed. A combination of bioinformatic and biochemical approaches allowed identification of phage promoters recognized by a host RNAP holoenzyme containing the σ70 factor. These promoters are located upstream of early phage genes. A combination of macroarray data, primer extension, and in vitro transcription analyses allowed identification of six promoters recognized by an RNAP holoenzyme containing gp36. These promoters are characterized by a single-consensus element tAATGTAtA and are located upstream of the middle and late phage genes. Curiously, gp79, an inhibitor of host and early phage transcription by σ70 holoenzyme, activated transcription by the gp36 holoenzyme in vitro.

Original languageEnglish (US)
Pages (from-to)389-399
Number of pages11
JournalJournal of molecular biology
Volume416
Issue number3
DOIs
StatePublished - Feb 24 2012

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Keywords

  • RNA polymerase
  • bacteriophage
  • genome
  • transcription regulation
  • σ factor

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