Transcriptional analysis of the Caulobacter 4.5 S RNA ffs gene and the physiological basis of an ffs mutant with a Ts phenotype

J Mol Biol. 1997 Oct 10;272(5):665-76. doi: 10.1006/jmbi.1997.1261.

Abstract

A temperature-sensitive (ts) mutation in the ffs gene, encoding 4.5 S RNA, gives rise to cell division and DNA replication defects in Caulobacter crescentus. The ffs gene is transcribed throughout the cell-cycle and is transcribed at similar rates in mutant (ffs36) and wild-type strains, but in the mutant the 4.5 S RNA is unstable leading to lower 4.5 S RNA levels. The ffs36 phenotype results from a single base change in one of the non-conserved stems of the mature RNA, and is completely rescued by a compensating mutation in the opposite strand, providing confirmation of the predicted secondary structure of the 4.5 S RNA. The Caulobacter ffs gene was shown to be functionally comparable to the Escherichia coli ffs gene by complementation. Comparison of the ffs36 strain to a ts secA strain of Caulobacter, also having cell-cycle and DNA replication phenotypes, showed that both exhibit a permanent induction of a heat shock response at the restrictive temperature. To explain the phenotype of both the secA and ffs36 strains, we propose that a cell-cycle checkpoint prevents further progression through the cell-cycle in response to increased intracellular levels of heat shock and misfolded proteins.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Caulobacter crescentus / cytology
  • Caulobacter crescentus / genetics*
  • Cell Cycle / physiology
  • Escherichia coli / chemistry
  • Escherichia coli / genetics
  • Gene Expression Regulation, Bacterial / genetics
  • Genes, Bacterial / genetics
  • Genetic Complementation Test
  • Heat-Shock Proteins / genetics
  • Heat-Shock Proteins / metabolism
  • Molecular Sequence Data
  • Mutation / genetics
  • Phenotype
  • Promoter Regions, Genetic / genetics
  • RNA, Bacterial / chemistry*
  • RNA, Bacterial / genetics*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sequence Analysis
  • Temperature
  • Transcription, Genetic / genetics

Substances

  • Heat-Shock Proteins
  • RNA, Bacterial
  • RNA, Messenger