Phosphoregulation of Nap1 Plays a Role in Septin Ring Dynamics and Morphogenesis in <named-content content-type="genus-species">Candida albicans</named-content>

ABSTRACT Nap1 has long been identified as a potential septin regulator in yeasts. However, its function and regulation remain poorly defined. Here, we report functional characterization of Nap1 in the human-pathogenic fungus Candida albicans. We find that deletion of NAP1 causes constitutive filamen...

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Autores principales:	Zhen-Xing Huang, Pan Zhao, Gui-Sheng Zeng, Yan-Ming Wang, Ian Sudbery, Yue Wang
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/9f578a99be8f47ef8311bd2650abca0f
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spelling	oai:doaj.org-article:9f578a99be8f47ef8311bd2650abca0f2021-11-15T15:45:10ZPhosphoregulation of Nap1 Plays a Role in Septin Ring Dynamics and Morphogenesis in <named-content content-type="genus-species">Candida albicans</named-content>10.1128/mBio.00915-132150-7511https://doaj.org/article/9f578a99be8f47ef8311bd2650abca0f2014-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00915-13https://doaj.org/toc/2150-7511ABSTRACT Nap1 has long been identified as a potential septin regulator in yeasts. However, its function and regulation remain poorly defined. Here, we report functional characterization of Nap1 in the human-pathogenic fungus Candida albicans. We find that deletion of NAP1 causes constitutive filamentous growth and changes of septin dynamics. We present evidence that Nap1’s cellular localization and function are regulated by phosphorylation. Phos-tag gel electrophoresis revealed that Nap1 phosphorylation is cell cycle dependent, exhibiting the lowest level around the time of bud emergence. Mass spectrometry identified 10 phosphoserine and phosphothreonine residues in a cluster near the N terminus, and mutation of these residues affected Nap1’s localization to the septin ring and cellular function. Nap1 phosphorylation involves two septin ring-associated kinases, Cla4 and Gin4, and its dephosphorylation occurs at the septin ring in a manner dependent on the phosphatases PP2A and Cdc14. Furthermore, the nap1Δ/Δ mutant and alleles carrying mutations of the phosphorylation sites exhibited greatly reduced virulence in a mouse model of systemic candidiasis. Together, our findings not only provide new mechanistic insights into Nap1’s function and regulation but also suggest the potential to target Nap1 in future therapeutic design. IMPORTANCE Septins are conserved filament-forming GTPases involved in a wide range of cellular events, such as cytokinesis, exocytosis, and morphogenesis. In Candida albicans, the most prevalent human fungal pathogen, septin functions are indispensable for its virulence. However, the molecular mechanisms by which septin structures are regulated are poorly understood. In this study, we deleted NAP1, a gene encoding a putative septin regulator, in C. albicans and found that cells lacking NAP1 showed abnormalities in morphology, invasive growth, and septin ring dynamics. We identified a conserved N-terminal phosphorylation cluster on Nap1 and demonstrated that phosphorylation at these sites regulates Nap1 localization and function. Importantly, deletion of NAP1 or mutation in the N-terminal phosphorylation cluster strongly reduced the virulence of C. albicans in a mouse model of systemic infection. Thus, this study not only provides mechanistic insights into septin regulation but also suggests Nap1 as a potential antifungal target.Zhen-Xing HuangPan ZhaoGui-Sheng ZengYan-Ming WangIan SudberyYue WangAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 1 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Zhen-Xing Huang Pan Zhao Gui-Sheng Zeng Yan-Ming Wang Ian Sudbery Yue Wang Phosphoregulation of Nap1 Plays a Role in Septin Ring Dynamics and Morphogenesis in <named-content content-type="genus-species">Candida albicans</named-content>
description	ABSTRACT Nap1 has long been identified as a potential septin regulator in yeasts. However, its function and regulation remain poorly defined. Here, we report functional characterization of Nap1 in the human-pathogenic fungus Candida albicans. We find that deletion of NAP1 causes constitutive filamentous growth and changes of septin dynamics. We present evidence that Nap1’s cellular localization and function are regulated by phosphorylation. Phos-tag gel electrophoresis revealed that Nap1 phosphorylation is cell cycle dependent, exhibiting the lowest level around the time of bud emergence. Mass spectrometry identified 10 phosphoserine and phosphothreonine residues in a cluster near the N terminus, and mutation of these residues affected Nap1’s localization to the septin ring and cellular function. Nap1 phosphorylation involves two septin ring-associated kinases, Cla4 and Gin4, and its dephosphorylation occurs at the septin ring in a manner dependent on the phosphatases PP2A and Cdc14. Furthermore, the nap1Δ/Δ mutant and alleles carrying mutations of the phosphorylation sites exhibited greatly reduced virulence in a mouse model of systemic candidiasis. Together, our findings not only provide new mechanistic insights into Nap1’s function and regulation but also suggest the potential to target Nap1 in future therapeutic design. IMPORTANCE Septins are conserved filament-forming GTPases involved in a wide range of cellular events, such as cytokinesis, exocytosis, and morphogenesis. In Candida albicans, the most prevalent human fungal pathogen, septin functions are indispensable for its virulence. However, the molecular mechanisms by which septin structures are regulated are poorly understood. In this study, we deleted NAP1, a gene encoding a putative septin regulator, in C. albicans and found that cells lacking NAP1 showed abnormalities in morphology, invasive growth, and septin ring dynamics. We identified a conserved N-terminal phosphorylation cluster on Nap1 and demonstrated that phosphorylation at these sites regulates Nap1 localization and function. Importantly, deletion of NAP1 or mutation in the N-terminal phosphorylation cluster strongly reduced the virulence of C. albicans in a mouse model of systemic infection. Thus, this study not only provides mechanistic insights into septin regulation but also suggests Nap1 as a potential antifungal target.
format	article
author	Zhen-Xing Huang Pan Zhao Gui-Sheng Zeng Yan-Ming Wang Ian Sudbery Yue Wang
author_facet	Zhen-Xing Huang Pan Zhao Gui-Sheng Zeng Yan-Ming Wang Ian Sudbery Yue Wang
author_sort	Zhen-Xing Huang
title	Phosphoregulation of Nap1 Plays a Role in Septin Ring Dynamics and Morphogenesis in <named-content content-type="genus-species">Candida albicans</named-content>
title_short	Phosphoregulation of Nap1 Plays a Role in Septin Ring Dynamics and Morphogenesis in <named-content content-type="genus-species">Candida albicans</named-content>
title_full	Phosphoregulation of Nap1 Plays a Role in Septin Ring Dynamics and Morphogenesis in <named-content content-type="genus-species">Candida albicans</named-content>
title_fullStr	Phosphoregulation of Nap1 Plays a Role in Septin Ring Dynamics and Morphogenesis in <named-content content-type="genus-species">Candida albicans</named-content>
title_full_unstemmed	Phosphoregulation of Nap1 Plays a Role in Septin Ring Dynamics and Morphogenesis in <named-content content-type="genus-species">Candida albicans</named-content>
title_sort	phosphoregulation of nap1 plays a role in septin ring dynamics and morphogenesis in <named-content content-type="genus-species">candida albicans</named-content>
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/9f578a99be8f47ef8311bd2650abca0f
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Phosphoregulation of Nap1 Plays a Role in Septin Ring Dynamics and Morphogenesis in <named-content content-type="genus-species">Candida albicans</named-content>

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