Super DksAs: Substitutions in DksA enhancing its effects on transcription initiation

Matthew D. Blankschien, Jeong Hyun Lee, Elicia D. Grace, Christopher W. Lennon, Jennifer A. Halliday, Wilma Ross, Richard L. Gourse, Christophe Herman

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

At specific times during bacterial growth, the transcription factor DksA and the unusual nucleotide regulator ppGpp work synergistically to inhibit some Escherichia coli promoters (e.g. rRNA promoters) and to stimulate others (e.g. promoters for amino-acid synthesis and transport). However, the mechanism of DksA action remains uncertain, in part because DksA does not function like conventional transcription factors. To gain insights into DksA function, we identified mutations in dksA that bypassed the requirement for ppGpp by selecting for growth of cells lacking ppGpp on minimal medium without amino acids. We show here that two substitutions in DksA, L15F and N88I, result in higher DksA activity both in vivo and in vitro, primarily by increasing the apparent affinity of DksA for RNA polymerase (RNAP). The mutant DksA proteins suggest potential roles for ppGpp in DksA function, identify potential surfaces on DksA crucial for RNAP binding, and provide tools for future studies to elucidate the mechanism of DksA action.

Original languageEnglish (US)
Pages (from-to)1720-1731
Number of pages12
JournalEMBO Journal
Volume28
Issue number12
DOIs
StatePublished - Jun 17 2009

Keywords

  • DksA
  • PpGpp
  • RNA polymerase secondary channel
  • Stringent response
  • Transcription factor

Fingerprint

Dive into the research topics of 'Super DksAs: Substitutions in DksA enhancing its effects on transcription initiation'. Together they form a unique fingerprint.

Cite this