Characterization of Escherichia coli cspE, whose product negatively regulates transcription of cspA, the gene for the major cold shock protein

Weonhye Bae, Sangita Phadtare, Konstantin Severinov, Masayori Inouye

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Abstract

Escherichia coli contains nine members of the CspA protein family from CspA to CspI. To elucidate the cellular function of CspE, we constructed a ΔcspE strain. CspE is highly produced at 37°C. The synthesis level of CspE transiently increased during the growth lag period after dilution of stationary-phase cells into the fresh medium at 37°C. This is consistent with the ΔcspE phenotype of the longer growth lag period after dilution. The protein synthesis patterns of the ΔcspE strain and the wild-type strain were compared using two-dimensional gel electrophoresis. In the ΔcspE strain, the synthesis of a number of proteins at 37°C was found to be altered and cspA was derepressed. The derepression of cspA in the ΔcspE strain was at the level of transcription in a promoter-independent fashion but was not caused by stabilization of the cspA mRNA, which was shown to be a major cause of CspA induction after cold shock. In vitro transcription assays demonstrated that both CspE and CspA enhanced transcription pause at the region immediately downstream of the cold box, a putative repressor binding site on the cspA mRNA. In a cell-free protein synthesis system using S-30 cell extracts, CspA production was specifically inhibited by the addition of CspE. These results indicate that CspE functions as a negative regulator for cspA expression at 37°C, probably by interacting with the transcription elongation complex at the cspA cold box region.

Original languageEnglish (US)
Pages (from-to)1429-1441
Number of pages13
JournalMolecular Microbiology
Volume31
Issue number5
DOIs
StatePublished - 1999

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

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