MAP Kinase Regulation of the <italic toggle="yes">Candida albicans</italic> Pheromone Pathway

ABSTRACT We investigated the relationships of the Cek1 and Cek2 mitogen-activated protein (MAP) kinases and the putative MAP kinase phosphatase Cpp1 in the mating process of Candida albicans. Mutants of the CPP1 gene are hyperresponsive to pheromone, generating large halos, high levels of projection...

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Autores principales:	Golnaz Rastghalam, Raha Parvizi Omran, Masoumeh Alizadeh, Debrah Fulton, Jaideep Mallick, Malcolm Whiteway
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Candida albicans GFP fusions MAP kinase phosphatases MAP kinases pheromone pathway Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/0b661eedf7d544c0a094d910f546dff3
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spelling	oai:doaj.org-article:0b661eedf7d544c0a094d910f546dff32021-11-15T15:22:05ZMAP Kinase Regulation of the <italic toggle="yes">Candida albicans</italic> Pheromone Pathway10.1128/mSphere.00598-182379-5042https://doaj.org/article/0b661eedf7d544c0a094d910f546dff32019-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00598-18https://doaj.org/toc/2379-5042ABSTRACT We investigated the relationships of the Cek1 and Cek2 mitogen-activated protein (MAP) kinases and the putative MAP kinase phosphatase Cpp1 in the mating process of Candida albicans. Mutants of the CPP1 gene are hyperresponsive to pheromone, generating large halos, high levels of projections, and an increase in pheromone-responsive gene expression. Mating-type-homozygous opaque cells that lack both kinases are sterile, consistent with previous observations, although several lines of evidence show that the two kinases do not simply provide redundant functions in the mating process. Loss of CEK1 reduces mating significantly, to about 0.3% of wild-type strains, and also reduces projection formation and pheromone-mediated gene expression. In contrast, loss of CEK2 has less of an effect, reducing mating to approximately one-third that of the wild-type strain and moderately reducing projection formation but having little influence on the induction of gene expression. However, loss of Cek2 function reduces adaptation to pheromone-mediated arrest. The mutation enhances pheromone response halos to a level similar to that of cpp1 mutants, although the cpp1 mutants are considerably more mating defective than the cek2 mutant. The double cek2 cpp1 mutant shows enhanced responsiveness relative to either single mutant in terms of gene expression and halo formation, suggesting the kinase and phosphatase roles in the adaptation process are independent. Analysis of protein phosphorylation shows that Cek1 undergoes pheromone-mediated phosphorylation of the activation loop, and this phosphorylation is enhanced in cells lacking either the Cpp1 phosphatase or the Cek2 kinase. In addition, Cek1-GFP shows enhanced nuclear localization in response to pheromone treatment. In contrast, Cek2 shows no evidence for pheromone-mediated phosphorylation or pheromone-mediated nuclear localization. Intriguingly, however, deletion of CPP1 enhances both the phosphorylation state and the nuclear localization of Cek2-GFP. Overall, these results identify a complex interaction among the MAP kinases and MAP kinase phosphatase that function in the C. albicans mating pathway. IMPORTANCE MAP kinases and their regulators are critical components of eukaryotic signaling pathways implicated in normal cell behavior as well as abnormal behaviors linked to diseases such as cancer. The mating pathway of the yeast Saccharomyces cerevisiae was central in establishing the MAP kinase paradigm. Here we investigate the mating pathway in a different ascomycete, the fungal pathogen C. albicans. In this dimorphic fungus MAP kinases are also implicated in the mating response, with two MAP kinases apparently playing redundant roles in the mating process. This work establishes that while some level of mating can occur in the presence of a single kinase, the Cek1 kinase is most important for mating, while the Cek2 kinase is involved in adaptation to signaling. While both kinases appear to be themselves regulated by dephosphorylation through the action of the Cpp1 phosphatase, this process appears important for mating only in the case of Cek1.Golnaz RastghalamRaha Parvizi OmranMasoumeh AlizadehDebrah FultonJaideep MallickMalcolm WhitewayAmerican Society for MicrobiologyarticleCandida albicansGFP fusionsMAP kinase phosphatasesMAP kinasespheromone pathwayMicrobiologyQR1-502ENmSphere, Vol 4, Iss 1 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Candida albicans GFP fusions MAP kinase phosphatases MAP kinases pheromone pathway Microbiology QR1-502
spellingShingle	Candida albicans GFP fusions MAP kinase phosphatases MAP kinases pheromone pathway Microbiology QR1-502 Golnaz Rastghalam Raha Parvizi Omran Masoumeh Alizadeh Debrah Fulton Jaideep Mallick Malcolm Whiteway MAP Kinase Regulation of the <italic toggle="yes">Candida albicans</italic> Pheromone Pathway
description	ABSTRACT We investigated the relationships of the Cek1 and Cek2 mitogen-activated protein (MAP) kinases and the putative MAP kinase phosphatase Cpp1 in the mating process of Candida albicans. Mutants of the CPP1 gene are hyperresponsive to pheromone, generating large halos, high levels of projections, and an increase in pheromone-responsive gene expression. Mating-type-homozygous opaque cells that lack both kinases are sterile, consistent with previous observations, although several lines of evidence show that the two kinases do not simply provide redundant functions in the mating process. Loss of CEK1 reduces mating significantly, to about 0.3% of wild-type strains, and also reduces projection formation and pheromone-mediated gene expression. In contrast, loss of CEK2 has less of an effect, reducing mating to approximately one-third that of the wild-type strain and moderately reducing projection formation but having little influence on the induction of gene expression. However, loss of Cek2 function reduces adaptation to pheromone-mediated arrest. The mutation enhances pheromone response halos to a level similar to that of cpp1 mutants, although the cpp1 mutants are considerably more mating defective than the cek2 mutant. The double cek2 cpp1 mutant shows enhanced responsiveness relative to either single mutant in terms of gene expression and halo formation, suggesting the kinase and phosphatase roles in the adaptation process are independent. Analysis of protein phosphorylation shows that Cek1 undergoes pheromone-mediated phosphorylation of the activation loop, and this phosphorylation is enhanced in cells lacking either the Cpp1 phosphatase or the Cek2 kinase. In addition, Cek1-GFP shows enhanced nuclear localization in response to pheromone treatment. In contrast, Cek2 shows no evidence for pheromone-mediated phosphorylation or pheromone-mediated nuclear localization. Intriguingly, however, deletion of CPP1 enhances both the phosphorylation state and the nuclear localization of Cek2-GFP. Overall, these results identify a complex interaction among the MAP kinases and MAP kinase phosphatase that function in the C. albicans mating pathway. IMPORTANCE MAP kinases and their regulators are critical components of eukaryotic signaling pathways implicated in normal cell behavior as well as abnormal behaviors linked to diseases such as cancer. The mating pathway of the yeast Saccharomyces cerevisiae was central in establishing the MAP kinase paradigm. Here we investigate the mating pathway in a different ascomycete, the fungal pathogen C. albicans. In this dimorphic fungus MAP kinases are also implicated in the mating response, with two MAP kinases apparently playing redundant roles in the mating process. This work establishes that while some level of mating can occur in the presence of a single kinase, the Cek1 kinase is most important for mating, while the Cek2 kinase is involved in adaptation to signaling. While both kinases appear to be themselves regulated by dephosphorylation through the action of the Cpp1 phosphatase, this process appears important for mating only in the case of Cek1.
format	article
author	Golnaz Rastghalam Raha Parvizi Omran Masoumeh Alizadeh Debrah Fulton Jaideep Mallick Malcolm Whiteway
author_facet	Golnaz Rastghalam Raha Parvizi Omran Masoumeh Alizadeh Debrah Fulton Jaideep Mallick Malcolm Whiteway
author_sort	Golnaz Rastghalam
title	MAP Kinase Regulation of the <italic toggle="yes">Candida albicans</italic> Pheromone Pathway
title_short	MAP Kinase Regulation of the <italic toggle="yes">Candida albicans</italic> Pheromone Pathway
title_full	MAP Kinase Regulation of the <italic toggle="yes">Candida albicans</italic> Pheromone Pathway
title_fullStr	MAP Kinase Regulation of the <italic toggle="yes">Candida albicans</italic> Pheromone Pathway
title_full_unstemmed	MAP Kinase Regulation of the <italic toggle="yes">Candida albicans</italic> Pheromone Pathway
title_sort	map kinase regulation of the <italic toggle="yes">candida albicans</italic> pheromone pathway
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/0b661eedf7d544c0a094d910f546dff3
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MAP Kinase Regulation of the <italic toggle="yes">Candida albicans</italic> Pheromone Pathway

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