1887

Abstract

() is a pathogenic fungus that infects humans and animals, and is responsible for the disease named sporotrichosis. The cell wall of this fungus has glycoproteins with a high content of mannose and rhamnose units, which are synthesized by endoplasmic reticulum- and Golgi-localized glycosyltransferases. Little is known about the enzymic machinery involved in the synthesis of these oligosaccharides in , or the genes encoding these activities. This is in part because of the lack of an available genome sequence for this organism. Using a partial genomic DNA library we identified Ss, whose predicted product has significant similarity to proteins encoded by members of the () / gene family. In order to biochemically characterize the putative enzyme, Ss was heterologously expressed in the methylotrophic yeast . Recombinant SsMnt1 showed Mn-dependent mannosyltransferase activity and the ability to recognize as acceptors α-methyl mannoside, mannose, ManGlcNAc oligosaccharide and a variety of mannobiosides. The characterization of the enzymic products generated by SsMnt1 revealed that the enzyme is an α1,2-mannosyltransferase that adds up to two mannose residues to the acceptor molecule. Functional complementation studies were performed in and mutants lacking members of the / gene family, demonstrating that SsMnt1 is involved in both the - and -linked glycosylation pathways, but not in phosphomannan elaboration.

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2012-09-01
2020-01-17
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