The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates

Detalhes bibliográficos
Autor(a) principal: Osborne, Morgan S
Data de Publicação: 2024
Outros Autores: Brehm, Joshua N, Olivença, Carmen, Cochran, Alicia M, Serrano, Mónica, Henriques, Adriano O, Sorg, Joseph A
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Texto Completo: http://hdl.handle.net/10362/182744
Resumo: Funding Information: We thank members of the Sorg laboratory for critical comments during the preparation of this manuscript. We also thank Joel Nott at The Protein Facility of Iowa State University for performing the Edman sequencing. This project was supported by awards 5R01AI116895 and 5R01AI172043 to J.A.S. from the National Institute of Allergy and Infectious Diseases. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIAID. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Publisher Copyright: © 2024 The Author(s). Molecular Microbiology published by John Wiley & Sons Ltd.
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spelling The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore SubstratesClostridioides difficilegerminationproteasesporeYabGMicrobiologyMolecular BiologyFunding Information: We thank members of the Sorg laboratory for critical comments during the preparation of this manuscript. We also thank Joel Nott at The Protein Facility of Iowa State University for performing the Edman sequencing. This project was supported by awards 5R01AI116895 and 5R01AI172043 to J.A.S. from the National Institute of Allergy and Infectious Diseases. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIAID. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Publisher Copyright: © 2024 The Author(s). Molecular Microbiology published by John Wiley & Sons Ltd.YabG is a sporulation-specific protease that is conserved among sporulating bacteria. Clostridioides difficile YabG processes the cortex destined proteins preproSleC into proSleC and CspBA to CspB and CspA. YabG also affects synthesis of spore coat/exosporium proteins CotA and CdeM. In prior work that identified CspA as the co-germinant receptor, mutations in yabG were found which altered the co-germinants required to initiate spore germination. To understand how these mutations in the yabG locus contribute to C. difficile spore germination, we introduced these mutations into an isogenic background. Spores derived from C. difficile yabGC207A (a catalytically inactive allele), C. difficile yabGA46D, C. difficile yabGG37E, and C. difficile yabGP153L strains germinated in response to taurocholic acid alone. Recombinantly expressed and purified preproSleC incubated with E. coli lysate expressing wild type YabG resulted in the removal of the presequence from preproSleC. Interestingly, only YabGA46D showed any activity toward purified preproSleC. Mutation of the YabG processing site in preproSleC (R119A) led to YabG shifting its processing to R115 or R112. Finally, changes in yabG expression under the mutant promoters were analyzed using a SNAP-tag and revealed expression differences at early and late stages of sporulation. Overall, our results support and expand upon the hypothesis that YabG is important for germination and spore assembly and, upon mutation of the processing site, can shift where it cleaves substrates.Instituto de Tecnologia Química e Biológica António Xavier (ITQB)Molecular, Structural and Cellular Microbiology (MOSTMICRO)RUNOsborne, Morgan SBrehm, Joshua NOlivença, CarmenCochran, Alicia MSerrano, MónicaHenriques, Adriano OSorg, Joseph A2025-05-07T21:19:13Z2024-09-112024-09-11T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/182744eng0950-382XPURE: 99908043https://doi.org/10.1111/mmi.15316info:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2025-05-19T01:39:21Zoai:run.unl.pt:10362/182744Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-29T07:13:46.613825Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates
title The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates
spellingShingle The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates
Osborne, Morgan S
Clostridioides difficile
germination
protease
spore
YabG
Microbiology
Molecular Biology
title_short The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates
title_full The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates
title_fullStr The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates
title_full_unstemmed The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates
title_sort The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates
author Osborne, Morgan S
author_facet Osborne, Morgan S
Brehm, Joshua N
Olivença, Carmen
Cochran, Alicia M
Serrano, Mónica
Henriques, Adriano O
Sorg, Joseph A
author_role author
author2 Brehm, Joshua N
Olivença, Carmen
Cochran, Alicia M
Serrano, Mónica
Henriques, Adriano O
Sorg, Joseph A
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
Molecular, Structural and Cellular Microbiology (MOSTMICRO)
RUN
dc.contributor.author.fl_str_mv Osborne, Morgan S
Brehm, Joshua N
Olivença, Carmen
Cochran, Alicia M
Serrano, Mónica
Henriques, Adriano O
Sorg, Joseph A
dc.subject.por.fl_str_mv Clostridioides difficile
germination
protease
spore
YabG
Microbiology
Molecular Biology
topic Clostridioides difficile
germination
protease
spore
YabG
Microbiology
Molecular Biology
description Funding Information: We thank members of the Sorg laboratory for critical comments during the preparation of this manuscript. We also thank Joel Nott at The Protein Facility of Iowa State University for performing the Edman sequencing. This project was supported by awards 5R01AI116895 and 5R01AI172043 to J.A.S. from the National Institute of Allergy and Infectious Diseases. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIAID. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Publisher Copyright: © 2024 The Author(s). Molecular Microbiology published by John Wiley & Sons Ltd.
publishDate 2024
dc.date.none.fl_str_mv 2024-09-11
2024-09-11T00:00:00Z
2025-05-07T21:19:13Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/182744
url http://hdl.handle.net/10362/182744
dc.language.iso.fl_str_mv eng
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dc.relation.none.fl_str_mv 0950-382X
PURE: 99908043
https://doi.org/10.1111/mmi.15316
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