Effect of sigma factor S (sigmaS) on the stability of penicillin-binding protein 3 (PBP3) of Escherichia colt K12

Abstract

The stability of penicillin-binding protein 3 (PBP3), a cell septum synthesizing protein, was analyzed at different incubation temperatures in three Escherichia coli K12 strains carrying a PBP3-overproducing plasmid. The stability of PBP3 was significantly reduced in stationary phase cells shifted to 42°C for 4 h, compared to samples incubated at 28 or 37°C. The half-life of PBP3 in the C600 strain was 60 min at 42°C, while samples incubated at 28 or 37°C had PBP3 half-lives greater than 4 h. Analysis of the PBP3 content in mutants deficient in rpoS (coding for the stationary phase sigma factor, sigmaS) and rpoH (coding for the heat shock sigma factor, sigma32) genes after shift to 42°C showed that stability of the protein was controlled by sigmaS but not by sigma32. These results suggest that control of the PBP3 levels in E. coli K12 is through a post-transcriptional mechanism regulated by the stationary phase regulon. We demonstrated that stability of PBP3 in E. coli K12 involves degradation of the protein. Moreover, we observed that incubation of cells at 42°C significantly reduces the stability of PBP3 in early stationary phase cells in a process controlled by sigmaS.

sigma factor S; penicillin-binding protein; Escherichia colt K12

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Effect of sigma factor S (sS) on the stability of penicillin-binding protein 3 (PBP3) of Escherichia colt K12^*

Rosane S. Machado; Douglas C. Camelo; Darcy F. de Almeida; Luis C.S. Ferreira

Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, CCS, 21949-900 Rio de Janeiro, RJ, Brasil. Tel.: [phone]; Fax: [phone]; E-mail: [e-mail]. Send correspondence to L.C.S.F.

ABSTRACT

The stability of penicillin-binding protein 3 (PBP3), a cell septum synthesizing protein, was analyzed at different incubation temperatures in three Escherichia coli K12 strains carrying a PBP3-overproducing plasmid. The stability of PBP3 was significantly reduced in stationary phase cells shifted to 42°C for 4 h, compared to samples incubated at 28 or 37°C. The half-life of PBP3 in the C600 strain was 60 min at 42°C, while samples incubated at 28 or 37°C had PBP3 half-lives greater than 4 h. Analysis of the PBP3 content in mutants deficient in rpoS (coding for the stationary phase sigma factor, sS) and rpoH (coding for the heat shock sigma factor, s32) genes after shift to 42°C showed that stability of the protein was controlled by sS but not by s32. These results suggest that control of the PBP3 levels in E. coli K12 is through a post-transcriptional mechanism regulated by the stationary phase regulon. We demonstrated that stability of PBP3 in E. coli K12 involves degradation of the protein. Moreover, we observed that incubation of cells at 42°C significantly reduces the stability of PBP3 in early stationary phase cells in a process controlled by sS.

Keywords: sigma factor S; penicillin-binding protein; Escherichia colt K12.

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*

This paper is dedicated to Prof. Carlos Chagas Filho, founder of the Instituto de Biofísica, on the occasion of its 50th anniversary.

Publication Dates

• Publication in this collection
  31 Oct 2006
• Date of issue
  1996