{"id":{"repo_id":"buffalo","oai_identifier":"oai:ubir.buffalo.edu:10477/86457"},"canonical_url":"https://search.dev.ndltd.org/etd/buffalo/oai:ubir.buffalo.edu:10477/86457","repository":{"repo_id":"buffalo","name":"Buffalo","base_url":"https://ubir.buffalo.edu/oai/request"},"display":{"title":"Candida albicans Glycosidases, Dfg5 and Dcw1, Regulate Cell Wall Chitin Synthesis","abstract":"M.S.","abstract_html":"M.S.","abstract_has_math":false,"creators":["Kim, Jaewon; 0000-0001-7952-0815"],"institution":"State University of New York at Buffalo","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Maddi, Abhiram","Oral Sciences"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025-02-21T17:22:37Z","date_published":"2025-02-21T17:22:37Z","updated_at":"2026-07-27T19:05:32Z","subjects":["dentistry"],"languages":["eng"],"rights":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/10477/86457","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Maddi, Abhiram","Oral Sciences"]},{"key":"dc:creator","label":"Author","values":["Kim, Jaewon; 0000-0001-7952-0815"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2025-02-21T17:22:37Z","2020"]},{"key":"dc:publisher","label":"Institution","values":["State University of New York at Buffalo"]},{"key":"dc:type","label":"Dc Type","values":["Text","Thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["dentistry"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/10477/86457"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["M.S.","Objectives: Candida albicans Dfg5 and Dcw1 are cell wall mannosidases with important functions and are ideal drug targets. Our past studies have shown that Dfg5 and Dcw1 not only function in cell wall biogenesis but they also regulate cell signaling via HOG MAPK signaling pathway. Hog1 plays a role in cell wall integrity via regulation of chitin synthases CHS1, CHS2, CHS3 and CHS8 in C. albicans. Our objective was to determine if Dfg5 and Dcw1 regulate chitin synthesis via HOG pathway. Methods: dfg5/dcw1 heterozygous (ES1, ES195) and conditional mutant (ES195+Methionine/Cysteine) strains were used. WT (SC5314) strain and DAY185 (parental) strains were used as negative controls and hog1 knock-out mutant as positive control. Fluorescence microscopy of calcofluor white (CFW) stained mutant and control strains was performed to determine characteristics of cell wall chitin. Quantification of chitin levels was performed by measuring CFW fluorescence of 100 cells per strain. Quantitative PCR analysis was performed to measure the relative expression of chitin synthase genes CHS1, CHS2, CHS3 and CHS8 in mutant and control strains. Strains were also treated with commercially available chitinase to determine cell separation using light microscopy and scanning electron microscopy (SEM) analysis. Results: Fluorescence microscopy showed significantly increased chitin accumulation in the mutants as compared to wild type. Chitin accumulation was observed mainly at the mother-bud neck. Additionally, incubation with chitinase led to cell separation in the mutants further confirming that the phenotype was a result of abnormal chitin accumulation. Chitin synthases CHS2, CHS3 and CHS8 were observed to be significantly upregulated in the ES195-conditional mutant at 6 h and 12 h as compared to the wild type. This upregulation was similar to hog1 knock-out mutant at 12 h for CHS2 and CHS8. A much higher increase in expression of CHS1 was observed in the hog1 knock-out mutant indicating that Hog1 may be required for regulation of CHS1. Conclusion: Cell wall glycosidases, Dfg5 and Dcw1, regulate chitin synthesis via expression of chitin synthases, which lead to differential chitin accumulation and a defect in cell separation.","**To request an accessible version of the file(s) associated with this item, contact library@buffalo.edu. Please include the item's persistent URL [http://hdl.handle.net/. . .] in your request.**"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Candida albicans Glycosidases, Dfg5 and Dcw1, Regulate Cell Wall Chitin Synthesis"]}]}],"canonical_facts":{"dc:contributor":["Maddi, Abhiram","Oral Sciences"],"dc:creator":["Kim, Jaewon; 0000-0001-7952-0815"],"dc:date":["2025-02-21T17:22:37Z","2020"],"dc:description":["M.S.","Objectives: Candida albicans Dfg5 and Dcw1 are cell wall mannosidases with important functions and are ideal drug targets. Our past studies have shown that Dfg5 and Dcw1 not only function in cell wall biogenesis but they also regulate cell signaling via HOG MAPK signaling pathway. Hog1 plays a role in cell wall integrity via regulation of chitin synthases CHS1, CHS2, CHS3 and CHS8 in C. albicans. Our objective was to determine if Dfg5 and Dcw1 regulate chitin synthesis via HOG pathway. Methods: dfg5/dcw1 heterozygous (ES1, ES195) and conditional mutant (ES195+Methionine/Cysteine) strains were used. WT (SC5314) strain and DAY185 (parental) strains were used as negative controls and hog1 knock-out mutant as positive control. Fluorescence microscopy of calcofluor white (CFW) stained mutant and control strains was performed to determine characteristics of cell wall chitin. Quantification of chitin levels was performed by measuring CFW fluorescence of 100 cells per strain. Quantitative PCR analysis was performed to measure the relative expression of chitin synthase genes CHS1, CHS2, CHS3 and CHS8 in mutant and control strains. Strains were also treated with commercially available chitinase to determine cell separation using light microscopy and scanning electron microscopy (SEM) analysis. Results: Fluorescence microscopy showed significantly increased chitin accumulation in the mutants as compared to wild type. Chitin accumulation was observed mainly at the mother-bud neck. Additionally, incubation with chitinase led to cell separation in the mutants further confirming that the phenotype was a result of abnormal chitin accumulation. Chitin synthases CHS2, CHS3 and CHS8 were observed to be significantly upregulated in the ES195-conditional mutant at 6 h and 12 h as compared to the wild type. This upregulation was similar to hog1 knock-out mutant at 12 h for CHS2 and CHS8. A much higher increase in expression of CHS1 was observed in the hog1 knock-out mutant indicating that Hog1 may be required for regulation of CHS1. Conclusion: Cell wall glycosidases, Dfg5 and Dcw1, regulate chitin synthesis via expression of chitin synthases, which lead to differential chitin accumulation and a defect in cell separation.","**To request an accessible version of the file(s) associated with this item, contact library@buffalo.edu. Please include the item's persistent URL [http://hdl.handle.net/. . .] in your request.**"],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/10477/86457"],"dc:language":["eng"],"dc:publisher":["State University of New York at Buffalo"],"dc:rights":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."],"dc:subject":["dentistry"],"dc:title":["Candida albicans Glycosidases, Dfg5 and Dcw1, Regulate Cell Wall Chitin Synthesis"],"dc:type":["Text","Thesis"]},"updated_at":"2026-07-27T19:05:32Z"}