Endopeptidase penicillin-binding proteins 4 and 7 play auxiliary roles in determining uniform morphology of Escherichia coli
TLDR
Overall cell shape may be determined by the existence or location of a specific type of peptide cross-link, with PBP 5 activity influencing how many cross- links are made and PBPs 4 and 7 acting as editing enzymes to remove inappropriate cross-links.Abstract:
The low-molecular-weight (LMW) penicillin-binding protein, PBP 5, plays a dominant role in determining the uniform cell shape of Escherichia coli. However, the physiological functions of six other LMW PBPs are unknown, even though the existence and enzymatic activities of four of these were established three decades ago. By applying fluorescence-activated cell sorting (FACS) to quantify the cellular dimensions of multiple PBP mutants, we found that the endopeptidases PBP 4 and PBP 7 also influence cell shape in concert with PBP 5. This is the first reported biological function for these two proteins. In addition, the combined loss of three dd-carboxypeptidases, PBPs 5 and 6 and DacD, also impaired cell shape. In contrast to previous reports based on visual inspection alone, FACS analysis revealed aberrant morphology in a mutant lacking only PBP 5, a phenotype not shared by any other strain lacking a single LMW PBP. PBP 5 removes the terminal d-alanine from pentapeptide side chains of muropeptide subunits, and pentapeptides act as donors for cross-linking adjacent side chains. As endopeptidases, PBPs 4 and 7 cleave cross-links in the cell wall. Therefore, overall cell shape may be determined by the existence or location of a specific type of peptide cross-link, with PBP 5 activity influencing how many cross-links are made and PBPs 4 and 7 acting as editing enzymes to remove inappropriate cross-links.read more
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Penicillin-binding protein 7/8 contributes to the survival of Acinetobacter baumannii in vitro and in vivo
Thomas A. Russo,Ulrike MacDonald,Janet M. Beanan,Ruth Olson,Ian J. MacDonald,Shauna L. Sauberan,L. Wayne Luke,L. Wayne Luke,Timothy C. Umland,Timothy C. Umland +9 more
TL;DR: It is demonstrated that PBP-7/8 contributes to the pathogenesis of A. baumannii and either directly or indirectly contributes toThe resistance of AB307-0294 to complement-mediated bactericidal activity.
Journal ArticleDOI
Septal and lateral wall localization of PBP5, the major D,D-carboxypeptidase of Escherichia coli, requires substrate recognition and membrane attachment
Lakshmi-Prasad Potluri,Aneta Karczmarek,Jolanda Verheul,André Piette,Jean-Marc Wilkin,Nadine Werth,Manuel Banzhaf,Waldemar Vollmer,Kevin D. Young,Martine Nguyen-Distèche,Tanneke den Blaauwen +10 more
TL;DR: The distribution of PBP5, the major D,D-carboxypeptidase in Escherichia coli, was mapped by immunolabelling and by visualization of GFP fusion proteins in wild-type cells and in mutants lacking one or more D-D-Carboxypepticases as mentioned in this paper.
Journal ArticleDOI
Dead‐end intermediates in the enterobacterial common antigen pathway induce morphological defects in Escherichia coli by competing for undecaprenyl phosphate
TL;DR: There is competition for a common pool of Und‐P, whose proper distribution to alternate metabolic pathways is required to maintain normal cell shape in E. coli, and the results argue that the buildup of ECA‐lipid II sequesters part of the pool ofundecaprenyl phosphate, which, in turn, adversely affects PG synthesis.
Journal ArticleDOI
Crystal structure of penicillin binding protein 4 (dacB) from Escherichia coli, both in the native form and covalently linked to various antibiotics
TL;DR: The crystal structure of penicillin binding protein 4 from Escherichia coli, which has both DD-endopeptidase and DD-carboxypeptid enzyme activity, is presented and the altered geometry of conserved active site residues compared with those of other PBPs suggests a possible cause for the slow deacylation rate of PBP4.
Journal ArticleDOI
Flow cytometry‐based enrichment for cell shape mutants identifies multiple genes that influence Helicobacter pylori morphology
Laura K. Sycuro,Chelsea S Rule,Chelsea S Rule,Timothy W. Petersen,Timna J. Wyckoff,Timna J. Wyckoff,Tate H. Sessler,Dilip B Nagarkar,Fakhra Khalid,Zachary Pincus,Jacoby Biboy,Waldemar Vollmer,Nina R. Salama,Nina R. Salama +13 more
TL;DR: FACS robustly detects perturbations of bacterial cell shape and identifies additional PG peptide modifications associated with helical cell shape in H. pylori, demonstrating its ability to identify shape‐determining genes.
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