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A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores
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 disea...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Academic Press
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8417463/ https://www.ncbi.nlm.nih.gov/pubmed/33940167 http://dx.doi.org/10.1016/j.anaerobe.2021.102379 |
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author | Alabdali, Yasir Adil Jabbar Oatley, Peter Kirk, Joseph A. Fagan, Robert P. |
author_facet | Alabdali, Yasir Adil Jabbar Oatley, Peter Kirk, Joseph A. Fagan, Robert P. |
author_sort | Alabdali, Yasir Adil Jabbar |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8417463 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Academic Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-84174632021-09-08 A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores Alabdali, Yasir Adil Jabbar Oatley, Peter Kirk, Joseph A. Fagan, Robert P. Anaerobe Clostridioides difficile (Including Epidemiology) 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. Academic Press 2021-08 /pmc/articles/PMC8417463/ /pubmed/33940167 http://dx.doi.org/10.1016/j.anaerobe.2021.102379 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Clostridioides difficile (Including Epidemiology) Alabdali, Yasir Adil Jabbar Oatley, Peter Kirk, Joseph A. Fagan, Robert P. A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores |
title | A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores |
title_full | A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores |
title_fullStr | A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores |
title_full_unstemmed | A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores |
title_short | A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores |
title_sort | cortex-specific penicillin-binding protein contributes to heat resistance in clostridioides difficile spores |
topic | Clostridioides difficile (Including Epidemiology) |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8417463/ https://www.ncbi.nlm.nih.gov/pubmed/33940167 http://dx.doi.org/10.1016/j.anaerobe.2021.102379 |
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