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Molecular characterization of adenylyl cyclase complex proteins using versatile protein-tagging plasmid systems in Cryptococcus neoformans
DC Field | Value | Language |
---|---|---|
dc.contributor.author | So, Yee-Seul | - |
dc.contributor.author | Yang, Dong-Hoon | - |
dc.contributor.author | Jung, Kwang-Woo | - |
dc.contributor.author | Huh, Won-Ki | - |
dc.contributor.author | Bahn, Yong-Sun | - |
dc.date.accessioned | 2023-04-19T08:43:47Z | - |
dc.date.available | 2023-04-19T08:43:47Z | - |
dc.date.created | 2018-01-10 | - |
dc.date.issued | 2017-02 | - |
dc.identifier.citation | Journal of Microbiology and Biotechnology, Vol.27 No.2, pp.357-364 | - |
dc.identifier.issn | 1017-7825 | - |
dc.identifier.uri | https://hdl.handle.net/10371/191116 | - |
dc.description.abstract | In this study, we aimed to generate a series of versatile tagging plasmids that can be used in diverse molecular biological studies of the fungal pathogen Cryptococcus neoformans. We constructed 12 plasmids that can be used to tag a protein of interest with a GFP, mCherry, 4xFLAG, or 6xHA, along with nourseothricin-, neomycin-, or hygromycin- resistant selection markers. Using this tagging plasmid set, we explored the adenylyl cyclase complex (ACC), consisting of adenylyl cyclase (Cac1) and its associated protein Aca1, in the cAMP-signaling pathway, which is critical for the pathogenicity of C. neoformans. We found that Cac1-mCherry and Aca1-GFP were mainly colocalized as punctate forms in the cell membrane and non-nuclear cellular organelles. We also demonstrated that Cac1 and Aca1 interacted in vivo by co-immunoprecipitation, using Cac1-6xHA and Aca1-4xFLAG tagging strains. Bimolecular fluorescence complementation further confirmed the in vivo interaction of Cac1 and Aca1 in live cells. Finally, protein pull-down experiments using aca1 Delta:: ACA1-GFP and aca1 Delta:: ACA1GFP cac1 Delta strains and comparative mass spectrometry analysis identified Cac1 and a number of other novel ACC-interacting proteins. Thus, this versatile tagging plasmid system will facilitate diverse mechanistic studies in C. neoformans and further our understanding of its biology. | - |
dc.language | 영어 | - |
dc.publisher | 한국미생물·생명공학회 | - |
dc.title | Molecular characterization of adenylyl cyclase complex proteins using versatile protein-tagging plasmid systems in Cryptococcus neoformans | - |
dc.type | Article | - |
dc.identifier.doi | 10.4014/jmb.1609.09036 | - |
dc.citation.journaltitle | Journal of Microbiology and Biotechnology | - |
dc.identifier.wosid | 000397168300018 | - |
dc.identifier.scopusid | 2-s2.0-85014416116 | - |
dc.citation.endpage | 364 | - |
dc.citation.number | 2 | - |
dc.citation.startpage | 357 | - |
dc.citation.volume | 27 | - |
dc.identifier.kciid | ART002201601 | - |
dc.description.isOpenAccess | N | - |
dc.contributor.affiliatedAuthor | Huh, Won-Ki | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.subject.keywordPlus | SIGNALING PATHWAYS | - |
dc.subject.keywordPlus | VIRULENCE | - |
dc.subject.keywordPlus | PATHOGENICITY | - |
dc.subject.keywordPlus | HOMEOSTASIS | - |
dc.subject.keywordPlus | ROLES | - |
dc.subject.keywordPlus | RAS | - |
dc.subject.keywordAuthor | Cyclic AMP | - |
dc.subject.keywordAuthor | adenylyl cyclase-associated protein | - |
dc.subject.keywordAuthor | Cac1 | - |
dc.subject.keywordAuthor | Aca1 | - |
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