title: Inactivation of CaMIT1 inhibits Candida albicans phospholipomannan beta-mannosylation, reduces virulence, and alters cell wall protein beta-mannosylation.
abstract: Studies on Candida albicans phospholipomannan have suggested a novel biosynthetic pathway for yeast glycosphingolipids. This pathway is thought to diverge from the usual pathway at the mannose-inositol-phospho-ceramide (MIPC) step. To confirm this hypothesis, a C. albicans gene homologue for the Saccharomyces cerevisiae SUR1 gene was identified and named MIT1 as it coded for GDP-mannose:inositol-phospho-ceramide mannose transferase. Two copies of this gene were disrupted. Western blots of cell extracts revealed that strain mit1Delta contained no PLM. Thin layer chromatography and mass spectrometry confirmed that mit1Delta did not synthesize MIPC, demonstrating a role of MIT1 in the mannosylation of C. albicans IPCs. As MIT1 disruption prevented downstream beta-1,2 mannosylation, mit1Delta represents a new C. albicans mutant affected in the expression of these specific virulence attributes, which act as adhesins/immunomodulators. mit1Delta was less virulent during both the acute and chronic phases of systemic infection in mice (75 and 50% reduction in mortality, respectively). In vitro, mit1Delta was not able to escape macrophage lysis through down-regulation of the ERK1/2 phosphorylation pathway previously shown to be triggered by PLM. Phenotypic analysis also revealed pleiotropic effects of MIT1 disruption. The most striking observation was a reduced beta-mannosylation of phosphopeptidomannan. Increased beta-mannosylation of mannoproteins was observed under growth conditions that prevented the association of beta-oligomannosides with phosphopeptidomannan, but not with PLM. This suggests that C. albicans has strong regulatory mechanisms associating beta-oligomannoses with different cell wall carrier molecules. These mechanisms and the impact of the different presentations of beta-oligomannoses on the host response need to be defined.
authors.affiliation: Laboratoire de Mycologie Fondamentale et Appliquée, Equipe Inserm E0360, Faculté de Médecine, Pôle Recherche, Place de Verdun, 59045 Lille Cedex, France.
authors.forename: Celine
authors.initials: C
authors.lastname: Mille
authors.forename: Guilhem
authors.initials: G
authors.lastname: Janbon
authors.forename: Florence
authors.initials: F
authors.lastname: Delplace
authors.forename: Stella
authors.initials: S
authors.lastname: Ibata-Ombetta
authors.forename: Claude
authors.initials: C
authors.lastname: Gaillardin
authors.forename: Gérard
authors.initials: G
authors.lastname: Strecker
authors.forename: Thierry
authors.initials: T
authors.lastname: Jouault
authors.forename: Pierre-André
authors.initials: PA
authors.lastname: Trinel
authors.forename: Daniel
authors.initials: D
authors.lastname: Poulain
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002176:Candida albicans / Q000166:cytology / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D002460:Cell Line; D048430:Cell Shape; D002473:Cell Wall / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D006017:Glycolipids / Q000737:chemistry / Q000378:metabolism*; D016695:Glycosyltransferases; D036341:Intercellular Signaling Peptides and Proteins; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology; D008358:Mannose / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010455:Peptides; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014166:Transferases / Q000235:genetics / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins; D006017:Glycolipids; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D010455:Peptides; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; C402142:mamba intestinal toxin 1; C400188:phospholipomannan; D014166:Transferases; D016695:Glycosyltransferases; C092031:SUR1 protein, S cerevisiae; D008358:Mannose; D002118:Calcium