Comprehensive Interactome Analysis for the Sole Adenylyl Cyclase Cyr1 of Candida albicans
ABSTRACT Cyr1, the sole adenylyl cyclase of the fungal pathogen Candida albicans, is a central component of the cAMP/protein kinase A signaling pathway that controls the yeast-to-hypha transition. Cyr1 is a multivalent sensor and integrator of various external and internal signals. To better underst...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2022-12-01
|
| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.03934-22 |
| _version_ | 1811194972415721472 |
|---|---|
| author | Guisheng Zeng Suat Peng Neo Li Mei Pang Jiaxin Gao Shu Chen Chong Jayantha Gunaratne Yue Wang |
| author_facet | Guisheng Zeng Suat Peng Neo Li Mei Pang Jiaxin Gao Shu Chen Chong Jayantha Gunaratne Yue Wang |
| author_sort | Guisheng Zeng |
| collection | DOAJ |
| description | ABSTRACT Cyr1, the sole adenylyl cyclase of the fungal pathogen Candida albicans, is a central component of the cAMP/protein kinase A signaling pathway that controls the yeast-to-hypha transition. Cyr1 is a multivalent sensor and integrator of various external and internal signals. To better understand how these signals are relayed to Cyr1 to regulate its activity, we sought to establish the interactome of Cyr1 by using stable isotope labeling by amino acids in cell culture (SILAC)-based quantitative proteomics to identify the proteins that coimmunoprecipitated with Cyr1. The method identified 36 proteins as candidates for authentic Cyr1-interacting partners, together with two known Cyr1-binding proteins, Cap1 and Act1. Fourteen identified proteins belonged to three functional groups, including actin regulation, cell wall components, and mitochondrial activities, that are known to play important roles in cell morphogenesis. To validate the proteomics data, we used biochemical and genetic methods to characterize two cell wall-related proteins, Mp65 and Sln1. First, coimmunoprecipitation confirmed their physical association with Cyr1. Second, deleting either MP65 or SLN1 resulted in severe defects in filamentation on serum plates. This study establishes the first Cyr1 interactome and uncovers a potential role for cell wall proteins in directly regulating Cyr1 activity to determine growth forms in C. albicans. IMPORTANCE A critical virulence trait of the human fungal pathogen Candida albicans is its ability to undergo the yeast-to-hypha transition in response to diverse environmental and cellular stimuli. Previous studies suggested that the sole adenylyl cyclase of C. albicans, Cyr1, is a multivalent signal sensor and integrator synthesizing cAMP to activate the downstream hypha-promoting events through the cAMP/protein kinase A pathway. To fully understand how Cyr1 senses and processes multiple stimuli to generate appropriate signal outputs, it was necessary to identify and characterize Cyr1-interacting partners. This study employed SILAC-based quantitative proteomic approaches and identified 36 Cyr1-associated proteins, many having functions associated with hyphal morphogenesis. Coimmunoprecipitation verified two cell surface proteins, Mp65 and Sln1. Furthermore, genetic and phenotypic analyses demonstrated the cAMP-dependent roles of these two proteins in determining hyphal growth. Our study establishes the first Cyr1 interactome and uncovers new Cyr1 regulators that mediate cell surface signals to influence the growth mode of C. albicans. |
| first_indexed | 2024-04-12T00:36:04Z |
| format | Article |
| id | doaj.art-51e168d903224b5a91d79f61e1058418 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-12T00:36:04Z |
| publishDate | 2022-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-51e168d903224b5a91d79f61e10584182022-12-22T03:55:09ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972022-12-0110610.1128/spectrum.03934-22Comprehensive Interactome Analysis for the Sole Adenylyl Cyclase Cyr1 of Candida albicansGuisheng Zeng0Suat Peng Neo1Li Mei Pang2Jiaxin Gao3Shu Chen Chong4Jayantha Gunaratne5Yue Wang6Infectious Diseases Labs, SingaporeQuantitative Proteomics Group, Institute of Molecular and Cell Biology, SingaporeInfectious Diseases Labs, SingaporeInfectious Diseases Labs, SingaporeInfectious Diseases Labs, SingaporeQuantitative Proteomics Group, Institute of Molecular and Cell Biology, SingaporeInfectious Diseases Labs, SingaporeABSTRACT Cyr1, the sole adenylyl cyclase of the fungal pathogen Candida albicans, is a central component of the cAMP/protein kinase A signaling pathway that controls the yeast-to-hypha transition. Cyr1 is a multivalent sensor and integrator of various external and internal signals. To better understand how these signals are relayed to Cyr1 to regulate its activity, we sought to establish the interactome of Cyr1 by using stable isotope labeling by amino acids in cell culture (SILAC)-based quantitative proteomics to identify the proteins that coimmunoprecipitated with Cyr1. The method identified 36 proteins as candidates for authentic Cyr1-interacting partners, together with two known Cyr1-binding proteins, Cap1 and Act1. Fourteen identified proteins belonged to three functional groups, including actin regulation, cell wall components, and mitochondrial activities, that are known to play important roles in cell morphogenesis. To validate the proteomics data, we used biochemical and genetic methods to characterize two cell wall-related proteins, Mp65 and Sln1. First, coimmunoprecipitation confirmed their physical association with Cyr1. Second, deleting either MP65 or SLN1 resulted in severe defects in filamentation on serum plates. This study establishes the first Cyr1 interactome and uncovers a potential role for cell wall proteins in directly regulating Cyr1 activity to determine growth forms in C. albicans. IMPORTANCE A critical virulence trait of the human fungal pathogen Candida albicans is its ability to undergo the yeast-to-hypha transition in response to diverse environmental and cellular stimuli. Previous studies suggested that the sole adenylyl cyclase of C. albicans, Cyr1, is a multivalent signal sensor and integrator synthesizing cAMP to activate the downstream hypha-promoting events through the cAMP/protein kinase A pathway. To fully understand how Cyr1 senses and processes multiple stimuli to generate appropriate signal outputs, it was necessary to identify and characterize Cyr1-interacting partners. This study employed SILAC-based quantitative proteomic approaches and identified 36 Cyr1-associated proteins, many having functions associated with hyphal morphogenesis. Coimmunoprecipitation verified two cell surface proteins, Mp65 and Sln1. Furthermore, genetic and phenotypic analyses demonstrated the cAMP-dependent roles of these two proteins in determining hyphal growth. Our study establishes the first Cyr1 interactome and uncovers new Cyr1 regulators that mediate cell surface signals to influence the growth mode of C. albicans.https://journals.asm.org/doi/10.1128/spectrum.03934-22Candida albicansCyr1SILACadenylyl cyclasemass spectrometry |
| spellingShingle | Guisheng Zeng Suat Peng Neo Li Mei Pang Jiaxin Gao Shu Chen Chong Jayantha Gunaratne Yue Wang Comprehensive Interactome Analysis for the Sole Adenylyl Cyclase Cyr1 of Candida albicans Microbiology Spectrum Candida albicans Cyr1 SILAC adenylyl cyclase mass spectrometry |
| title | Comprehensive Interactome Analysis for the Sole Adenylyl Cyclase Cyr1 of Candida albicans |
| title_full | Comprehensive Interactome Analysis for the Sole Adenylyl Cyclase Cyr1 of Candida albicans |
| title_fullStr | Comprehensive Interactome Analysis for the Sole Adenylyl Cyclase Cyr1 of Candida albicans |
| title_full_unstemmed | Comprehensive Interactome Analysis for the Sole Adenylyl Cyclase Cyr1 of Candida albicans |
| title_short | Comprehensive Interactome Analysis for the Sole Adenylyl Cyclase Cyr1 of Candida albicans |
| title_sort | comprehensive interactome analysis for the sole adenylyl cyclase cyr1 of candida albicans |
| topic | Candida albicans Cyr1 SILAC adenylyl cyclase mass spectrometry |
| url | https://journals.asm.org/doi/10.1128/spectrum.03934-22 |
| work_keys_str_mv | AT guishengzeng comprehensiveinteractomeanalysisforthesoleadenylylcyclasecyr1ofcandidaalbicans AT suatpengneo comprehensiveinteractomeanalysisforthesoleadenylylcyclasecyr1ofcandidaalbicans AT limeipang comprehensiveinteractomeanalysisforthesoleadenylylcyclasecyr1ofcandidaalbicans AT jiaxingao comprehensiveinteractomeanalysisforthesoleadenylylcyclasecyr1ofcandidaalbicans AT shuchenchong comprehensiveinteractomeanalysisforthesoleadenylylcyclasecyr1ofcandidaalbicans AT jayanthagunaratne comprehensiveinteractomeanalysisforthesoleadenylylcyclasecyr1ofcandidaalbicans AT yuewang comprehensiveinteractomeanalysisforthesoleadenylylcyclasecyr1ofcandidaalbicans |