A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores.

Autor: Alabdali YAJ; Florey Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, UK., Oatley P; Florey Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, UK., Kirk JA; Florey Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, UK., Fagan RP; Florey Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, UK. Electronic address: r.fagan@sheffield.ac.uk.
Jazyk: angličtina
Zdroj: Anaerobe [Anaerobe] 2021 Aug; Vol. 70, pp. 102379. Date of Electronic Publication: 2021 Apr 30.
DOI: 10.1016/j.anaerobe.2021.102379
Abstrakt: Background: Sporulation is a complex cell differentiation programme shared by many members of the Firmicutes, the end result of which is a highly resistant, metabolically inert spore that can survive harsh environmental insults. Clostridioides difficile spores are essential for transmission of disease and are also required for recurrent infection. However, the molecular basis of sporulation is poorly understood, despite parallels with the well-studied Bacillus subtilis system. The spore envelope consists of multiple protective layers, one of which is a specialised layer of peptidoglycan, called the cortex, that is essential for the resistant properties of the spore. We set out to identify the enzymes required for synthesis of cortex peptidoglycan in C. difficile.
Methods: Bioinformatic analysis of the C. difficile genome to identify putative homologues of Bacillus subtilis spoVD was combined with directed mutagenesis and microscopy to identify and characterise cortex-specific PBP activity.
Results: Deletion of CDR20291_2544 (SpoVD Cd ) abrogated spore formation and this phenotype was completely restored by complementation in cis. Analysis of SpoVD Cd revealed a three domain structure, consisting of dimerization, transpeptidase and PASTA domains, very similar to B. subtilis SpoVD. Complementation with SpoVD Cd domain mutants demonstrated that the PASTA domain was dispensable for formation of morphologically normal spores. SpoVD Cd was also seen to localise to the developing spore by super-resolution confocal microscopy.
Conclusions: We have identified and characterised a cortex specific PBP in C. difficile. This is the first characterisation of a cortex-specific PBP in C. difficile and begins the process of unravelling cortex biogenesis in this important pathogen.
Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
(Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
Databáze: MEDLINE
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