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Overproduction of Inactive Variants of the Murein Synthase PBP1B Causes Lysis in Escherichia coli

By Ute Meisel, Joachim-Volker Höltje and Waldemar Vollmer

Abstract

Penicillin-binding protein 1B (PBP1B) of Escherichia coli is a bifunctional murein synthase containing both a transpeptidase domain and a transglycosylase domain. The protein is present in three forms (α, β, and γ) which differ in the length of their N-terminal cytoplasmic region. Expression plasmids allowing the production of native PBP1B or of PBP1B variants with an inactive transpeptidase or transglycosylase domain or both were constructed. The inactive domains contained a single amino acid exchange in an essential active-site residue. Overproduction of the inactive PBP1B variants, but not of the active proteins, caused lysis of wild-type cells. The cells became tolerant to lysis by inactive PBP1B at a pH of 5.0, which is similar to the known tolerance for penicillin-induced lysis under acid pH conditions. Lysis was also reduced in mutant strains lacking several murein hydrolases. In particular, a strain devoid of activity of all known lytic transglycosylases was virtually tolerant, indicating that mainly the lytic transglycosylases are responsible for the observed lysis effect. A possible structural interaction between PBP1B and murein hydrolases in vivo by the formation of a multienzyme complex is discussed

Topics: Microbial Cell Biology
Publisher: American Society for Microbiology
Year: 2003
DOI identifier: 10.1128/JB.185.18.5342-5348.2003
OAI identifier: oai:pubmedcentral.nih.gov:193747
Provided by: PubMed Central
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