CsrA regulates translation of the Escherichia coli carbon starvation gene, cstA, by blocking ribosome access to the cstA transcript

Ashok K. Dubey, Carol Sue Baker, Kazushi Suzuki, A. Daniel Jones, Pallavi Pandit, Tony Romeo, Paul Lee Babitzke

Research output: Contribution to journalArticle

126 Citations (Scopus)

Abstract

CsrA is a global regulator that binds to two sites in the glgCAP leader transcript, thereby blocking ribosome access to the glgC Shine-Dalgarno sequence. The upstream CsrA binding site (GCACACGGAU) was used to search the Escherichia coli genomic sequence for other genes that might be regulated by CsrA. cstA contained an exact match that overlapped its Shine-Dalgarno sequence. cstA was previously shown to be induced by carbon starvation and to encode a peptide transporter. Expression of a cstA′-′lacZ translational fusion in wild-type and csrA mutant strains was examined. Expression levels in the csrA mutant were approximately twofold higher when cells were grown in Luria broth (LB) and 5- to 10-fold higher when LB was supplemented with glucose. It was previously shown that cstA is regulated by the cyclic AMP (cAMP)-cAMP receptor protein complex and transcribed by Eσ70. We investigated the influence of σS on cstA expression and found that a σS deficiency resulted in a threefold increase in cstA expression in wild-type and csrA mutant strains; however, CsrA-dependent regulation was retained. The mechanism of CsrA-mediated cstA regulation was also examined in vitro. Cross-linking studies demonstrated that CsrA is a homodimer. Gel mobility shift results showed that CsrA binds specifically to cstA RNA, while coupled-transcription-translation and toeprint studies demonstrated that CsrA regulates CstA synthesis by inhibiting ribosome binding to cstA transcripts. RNA footprint and boundary analyses revealed three or four CsrA binding sites, one of which overlaps the cstA Shine-Dalgarno sequence, as predicted. These results establish that CsrA regulates translation of cstA by sterically interfering with ribosome binding.

Original languageEnglish (US)
Pages (from-to)4450-4460
Number of pages11
JournalJournal of bacteriology
Volume185
Issue number15
DOIs
StatePublished - Aug 1 2003

Fingerprint

Starvation
Ribosomes
Carbon
Escherichia coli
Binding Sites
Cyclic AMP Receptor Protein
RNA
Genes
Cyclic AMP
Gels
Glucose

All Science Journal Classification (ASJC) codes

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Dubey, Ashok K. ; Baker, Carol Sue ; Suzuki, Kazushi ; Jones, A. Daniel ; Pandit, Pallavi ; Romeo, Tony ; Babitzke, Paul Lee. / CsrA regulates translation of the Escherichia coli carbon starvation gene, cstA, by blocking ribosome access to the cstA transcript. In: Journal of bacteriology. 2003 ; Vol. 185, No. 15. pp. 4450-4460.
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CsrA regulates translation of the Escherichia coli carbon starvation gene, cstA, by blocking ribosome access to the cstA transcript. / Dubey, Ashok K.; Baker, Carol Sue; Suzuki, Kazushi; Jones, A. Daniel; Pandit, Pallavi; Romeo, Tony; Babitzke, Paul Lee.

In: Journal of bacteriology, Vol. 185, No. 15, 01.08.2003, p. 4450-4460.

Research output: Contribution to journalArticle

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