Publicação
The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates
| Resumo: | 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. |
|---|---|
| Autores principais: | Osborne, Morgan S |
| Outros Autores: | Brehm, Joshua N; Olivença, Carmen; Cochran, Alicia M; Serrano, Mónica; Henriques, Adriano O; Sorg, Joseph A |
| Assunto: | Clostridioides difficile germination protease spore YabG Microbiology Molecular Biology |
| Ano: | 2024 |
| País: | Portugal |
| Tipo de documento: | artigo |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade Nova de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Institucional da UNL |
| _version_ | 1868983547801370624 |
|---|---|
| author | Osborne, Morgan S |
| 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 |
| author_facet | Osborne, Morgan S Brehm, Joshua N Olivença, Carmen Cochran, Alicia M Serrano, Mónica Henriques, Adriano O Sorg, Joseph A |
| author_role | author |
| contributor_name_str_mv | Instituto de Tecnologia Química e Biológica António Xavier (ITQB) Molecular, Structural and Cellular Microbiology (MOSTMICRO) Wiley RUN |
| country_str | PT |
| creators_json_txt | [{\"Person.name\":\"Osborne, Morgan S\"},{\"Person.name\":\"Brehm, Joshua N\"},{\"Person.name\":\"Olivença, Carmen\"},{\"Person.name\":\"Cochran, Alicia M\"},{\"Person.name\":\"Serrano, Mónica\"},{\"Person.name\":\"Henriques, Adriano O\"},{\"Person.name\":\"Sorg, Joseph A\"}] |
| datacite.contributors.contributor.contributorName.fl_str_mv | Instituto de Tecnologia Química e Biológica António Xavier (ITQB) Molecular, Structural and Cellular Microbiology (MOSTMICRO) Wiley RUN |
| datacite.creators.creator.creatorName.fl_str_mv | Osborne, Morgan S Brehm, Joshua N Olivença, Carmen Cochran, Alicia M Serrano, Mónica Henriques, Adriano O Sorg, Joseph A |
| datacite.date.Accepted.fl_str_mv | 2024-09-11T00:00:00Z |
| datacite.date.available.fl_str_mv | 2025-05-07T21:19:13Z |
| datacite.date.embargoed.fl_str_mv | 2025-05-07T21:19:13Z |
| datacite.rights.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| datacite.subjects.subject.fl_str_mv | Clostridioides difficile germination protease spore YabG Microbiology Molecular Biology |
| datacite.titles.title.fl_str_mv | The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates |
| dc.contributor.none.fl_str_mv | Instituto de Tecnologia Química e Biológica António Xavier (ITQB) Molecular, Structural and Cellular Microbiology (MOSTMICRO) Wiley RUN |
| dc.creator.none.fl_str_mv | Osborne, Morgan S Brehm, Joshua N Olivença, Carmen Cochran, Alicia M Serrano, Mónica Henriques, Adriano O Sorg, Joseph A |
| dc.date.Accepted.fl_str_mv | 2024-09-11T00:00:00Z |
| dc.date.available.fl_str_mv | 2025-05-07T21:19:13Z |
| dc.date.embargoed.fl_str_mv | 2025-05-07T21:19:13Z |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | http://hdl.handle.net/10362/182744 |
| dc.language.none.fl_str_mv | eng |
| dc.rights.none.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| dc.subject.none.fl_str_mv | Clostridioides difficile germination protease spore YabG Microbiology Molecular Biology |
| dc.title.fl_str_mv | The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates |
| dc.type.none.fl_str_mv | http://purl.org/coar/resource_type/c_6501 |
| description | 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. |
| dirty | 0 |
| eu_rights_str_mv | openAccess |
| format | article |
| fulltext.url.fl_str_mv | https://run.unl.pt/bitstreams/10d3d51c-46bf-4a36-8b15-a4e259873993/download |
| id | run_a2ed8dccbff24d769bbfa09c22a7cd03 |
| identifier.url.fl_str_mv | http://hdl.handle.net/10362/182744 |
| inst_facet_str | urn:organizationAcronym:unl{{{_:::_}}}Universidade Nova de Lisboa |
| instacron_str | unl |
| institution | Universidade Nova de Lisboa |
| instname_str | Universidade Nova de Lisboa |
| language | eng |
| network_acronym_str | run |
| network_name_str | Repositório Institucional da UNL |
| oai_identifier_str | oai:run.unl.pt:10362/182744 |
| organization_str_mv | urn:organizationAcronym:unl |
| person_str_mv | Osborne, Morgan S Brehm, Joshua N Olivença, Carmen Cochran, Alicia M Serrano, Mónica Henriques, Adriano O Sorg, Joseph A |
| publishDate | 2024 |
| repo_facet_str | urn:repositoryAcronym:run{{{_:::_}}}Repositório Institucional da UNL |
| reponame_str | Repositório Institucional da UNL |
| repository_id_str | urn:repositoryAcronym:run |
| service_str_mv | urn:repositoryAcronym:run |
| spelling | engenYabG 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.application/pdfenThe Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore SubstratesOsborne, Morgan SBrehm, Joshua NOlivença, CarmenCochran, Alicia MSerrano, MónicaHenriques, Adriano OSorg, Joseph AInstituto de Tecnologia Química e Biológica António Xavier (ITQB)Molecular, Structural and Cellular Microbiology (MOSTMICRO)WileyHostingInstitutionOrganizationalRUNe-mailmailto:run@unl.ptrun@unl.ptISSNIsPartOf0950-382XURNIsPartOfPURE: 99908043URNIsPartOfPURE UUID: eac966ca-926d-407a-9fe6-c42cf5111933URNIsPartOfScopus: 85203535325DOIIsPartOf10.1111/mmi.153162025-05-07T21:19:13Z2024-09-112024-09-11T00:00:00ZHandlehttp://hdl.handle.net/10362/182744http://purl.org/coar/access_right/c_abf2open accessClostridioides difficilegerminationproteasesporeYabGMicrobiologyMolecular Biology16198653 bytesliteraturehttp://purl.org/coar/resource_type/c_6501journal articlehttp://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://run.unl.pt/bitstreams/10d3d51c-46bf-4a36-8b15-a4e259873993/download |
| 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 |
| status | SINGLETON |
| subject.fl_str_mv | Clostridioides difficile germination protease spore YabG Microbiology Molecular Biology |
| title | 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_short | 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 |
| topic | Clostridioides difficile germination protease spore YabG Microbiology Molecular Biology |
| topic_facet | Clostridioides difficile germination protease spore YabG Microbiology Molecular Biology |
| url | http://hdl.handle.net/10362/182744 |
| visible | 1 |