Characterization of HscC (Hsc62), homologue of Hsp70 in Escherichia coli: over-expression of HscC modulates the activity of house keeping sigma factor sigma70

Genes Cells. 2002 Jun;7(6):553-66. doi: 10.1046/j.1365-2443.2002.00545.x.

Abstract

Background: HscC, the third member of the Hsp70 family in Escherichia coli, shares 33% identity with the other two homologues, DnaK and HscA, and displays ATPase activity. Genetic and biochemical evidence indicates that the DnaK-DnaJ chaperone system interacts with sigma32 and is involved in the negative regulation of the heat shock response. Although HscC is a highly conserved protein in the Hsp70 family, its function is still unknown.

Results: We observed that the over-expression of HscC caused severe growth inhibition. To explore this effect, we performed primer extension analysis and a beta-galactosidase assay and found that HscC reduced the sigma70-dependent promoter activity. An in vitro transcription assay revealed that HscC inhibited sigma70-dependent transcription. In addition, the co-purification analysis showed that sigma70 co-eluted with HscC.

Conclusion: These results indicate that HscC forms a complex with sigma70 and may function as its negative modulator.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA-Directed RNA Polymerases / metabolism*
  • Escherichia coli / genetics
  • Escherichia coli / growth & development
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Gene Expression Regulation, Bacterial
  • HSP70 Heat-Shock Proteins / genetics
  • HSP70 Heat-Shock Proteins / metabolism*
  • Promoter Regions, Genetic
  • Sigma Factor / metabolism*

Substances

  • Escherichia coli Proteins
  • HSP70 Heat-Shock Proteins
  • Sigma Factor
  • hscC protein, E coli
  • RNA polymerase sigma 70
  • DNA-Directed RNA Polymerases
  • dnaK protein, E coli