Dataset Information

The metacaspase Mca1p restricts O-glycosylation during farnesol-induced apoptosis in Candida albicans

ABSTRACT: Protein glycolysation is an essential posttranslational modification in eukaryotic cells. In pathogenic yeasts, it is involved in a large number of biological processes, such as protein folding quality control, cell viability and host/pathogen relationships. A link between protein glycosylation and apoptosis was established by the analysis of the phenotypes of oligosaccharyltransferase mutants in budding yeast. However, little is known about the contribution of glycosylation modifications to the adaptive response to apoptosis inducers. The cysteine protease metacaspase Mca1p plays a key role in the apoptotic response in Candida albicans triggered by the quorum sensing molecule farnesol. We subjected wild-type and mca1-deletion strains to farnesol stress and then studied the early phase of apoptosis release in quantitative glycoproteomics and glycomics experiments on cell-free extracts essentially devoid of cell walls. We identified and characterized 62 new glycosylated peptides with their glycan composition: 17 N-glycosylated, 45 O-glycosylated, and 81 additional sites of N-glycosylation. The glycosylated proteins were predicted as located mostly in the “membrane” and “cell wall” compartments, but they were also related to the “nucleus” and the “mitochondrial” compartments. They were found to be involved in the control of protein folding, cell wall integrity and cell cycle regulation. We showed a general increase in the O-glycosylation of proteins in the mca1 deletion strain after farnesol challenge. We identified 44 new putative protein substrates of the metacaspase in the glycoprotein fraction enriched on concanavalin A. Most of these substrates are involved in protein folding or protein resolubilization and in mitochondrial functions. We show here that key Mca1p substrates, such as Cdc48p or Ssb1p, involved in degrading misfolded glycoproteins and in the protein quality control system, are themselves differentially glycosylated. We found putative substrates, such as Bgl2p, Srb1p or Ugp1p, that are involved in the biogenesis of glycans. We validated as a substrate of Mca1p the endo-beta-1,3-glucanase Bgl2p which is involved in the incorporation of newly synthetized mannoproteins in the cell wall. Our findings highlight a new role of the metacaspase in amplifying cell death processes by affecting several critical protein quality control systems through the alteration of the protein glycosylation machinery.

INSTRUMENT(S): Orbitrap Fusion ETD

ORGANISM(S): Candida Albicans (yeast)

TISSUE(S): Cell Culture, Vegetative Cell (sensu Fungi)

SUBMITTER: Thibaut Léger

LAB HEAD: Jean-Michel Camadro

PROVIDER: PXD003677 | Pride | 2016-05-02

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	1450005-ConA-proteins-MCA1-1.raw	Raw
	1450005-ConA-proteins-MCA1-2.raw	Raw
	1450005-ConA-proteins-MCA1-3.raw	Raw
	1450005-ConA-proteins-MCA1F-1.raw	Raw
	1450005-ConA-proteins-MCA1F-2.raw	Raw

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The Metacaspase (Mca1p) Restricts O-glycosylation During Farnesol-induced Apoptosis in Candida albicans.

Léger Thibaut T Garcia Camille C Camadro Jean-Michel JM

Molecular & cellular proteomics : MCP 20160428 7

Protein glycolysation is an essential posttranslational modification in eukaryotic cells. In pathogenic yeasts, it is involved in a large number of biological processes, such as protein folding quality control, cell viability and host/pathogen relationships. A link between protein glycosylation and apoptosis was established by the analysis of the phenotypes of oligosaccharyltransferase mutants in budding yeast. However, little is known about the contribution of glycosylation modifications to the ...[more]

PMID: 27125826

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