MNN5 encodes an iron-regulated alpha-1,2-mannosyltransferase important for protein glycosylation, cell wall integrity, morphogenesis, and virulence in Candida albicans.
The cell walls of microbial pathogens mediate physical interactions with host cells and hence play a key role in infection. Mannosyltransferases have been shown to determine the cell wall properties and virulence of the pathogenic fungus Candida albicans. We previously identified a C. albicans alpha-1,2-mannosyltransferase, Mnn5, for its novel ability to enhance iron usage in Saccharomyces cerevisiae. Here we have studied the enzymatic properties of purified Mnn5 and characterized its function in its natural host. Mnn5 catalyzes the transfer of mannose to both alpha-1,2- and alpha-1,6-mannobiose, and this activity requires Mn2+ as a cofactor and is regulated by the Fe2+ concentration. An mnn5Delta mutant showed a lowered ability to extend O-linked, and possibly also N-linked, mannans, hypersensitivity to cell wall-damaging agents, and a reduction of cell wall mannosylphosphate content, phenotypes typical of many fungal mannosyltransferase mutants. The mnn5Delta mutant also exhibited some unique defects, such as impaired hyphal growth on solid media and attenuated virulence in mice. An unanticipated phenotype was the mnn5Delta mutant's resistance to killing by the iron-chelating protein lactoferrin, rendering it the first protein found that mediates lactoferrin killing of C. albicans. In summary, MNN5 deletion impairs a wide range of cellular events, most likely due to its broad substrate specificity. Of particular interest was the observed role of iron in regulating the enzymatic activity, suggesting an underlying relationship between Mnn5 activity and cellular iron homeostasis.
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Singapore 138673, Singapore.
Chen
C
Bai
Xiao-Li
XL
Xu
Fong-Yee
FY
Chan
Raymond Teck Ho
RT
Lee
Yue
Y
Wang
D000423:Alcian Blue; D000818:Animals; D002176:Candida albicans / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development / Q000472:pathogenicity*; D002473:Cell Wall / Q000378:metabolism*; D017353:Gene Deletion; D015870:Gene Expression; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D005800:Genes, Fungal / Q000235:genetics; D006031:Glycosylation; D025301:Hyphae / Q000166:cytology; D007501:Iron / Q000494:pharmacology*; D007781:Lactoferrin / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008364:Mannosyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D051379:Mice; D009024:Morphogenesis; D012441:Saccharomyces cerevisiae; D014774:Virulence
D008345:Manganese; D007501:Iron; D008364:Mannosyltransferases; D007781:Lactoferrin; D000423:Alcian Blue