title: The putative alpha-1,2-mannosyltransferase AfMnt1 of the opportunistic fungal pathogen Aspergillus fumigatus is required for cell wall stability and full virulence.
abstract: Proteins entering the eukaryotic secretory pathway commonly are glycosylated. Important steps in this posttranslational modification are carried out by mannosyltransferases. In this study, we investigated the putative alpha-1,2-mannosyltransferase AfMnt1 of the human pathogenic mold Aspergillus fumigatus. AfMnt1 belongs to a family of enzymes that comprises nine members in Saccharomyces cerevisiae but only three in A. fumigatus. A Deltaafmnt1 mutant is viable and grows normally at 37 degrees C, but its hyphal cell wall appears to be thinner than that of the parental strain. The lack of AfMnt1 leads to a higher sensitivity to calcofluor white and Congo red but not to sodium dodecyl sulfate. The growth of the mutant is abrogated at 48 degrees C but can be restored by osmotic stabilization. The resulting colonies remain white due to a defect in the formation of conidia. Electron and immunofluorescence microscopy further revealed that the observed growth defect of the mutant at 48 degrees C can be attributed to cell wall instability resulting in leakage at the hyphal tips. Using a red fluorescence fusion protein, we localized AfMnt1 in compact, brefeldin A-sensitive organelles that most likely represent fungal Golgi equivalents. The tumor necrosis factor alpha response of murine macrophages to hyphae was not affected by the lack of the afmnt1 gene, but the corresponding mutant was attenuated in a mouse model of infection. This and the increased sensitivity of the Deltaafmnt1 mutant to azoles, antifungal agents that currently are used to treat Aspergillus infections, suggest that alpha-1,2-mannosyltransferases are interesting targets for novel antifungal drugs.
authors: Max-von-Pettenkofer-Institute, Ludwig-Maximilians-Universität, Munich, Germany.
authors: Johannes
authors: J
authors: Wagener
authors: Bernd
authors: B
authors: Echtenacher
authors: Manfred
authors: M
authors: Rohde
authors: Andrea
authors: A
authors: Kotz
authors: Sven
authors: S
authors: Krappmann
authors: Jürgen
authors: J
authors: Heesemann
authors: Frank
authors: F
authors: Ebel
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D001228:Aspergillosis / Q000276:immunology / Q000382:microbiology*; D001232:Aspergillus fumigatus / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D002473:Cell Wall / Q000201:enzymology* / Q000235:genetics; D016207:Cytokines / Q000276:immunology; D005260:Female; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D025301:Hyphae / Q000201:enzymology / Q000235:genetics; D008264:Macrophages / Q000276:immunology / Q000382:microbiology; D008364:Mannosyltransferases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011995:Recombination, Genetic; D017384:Sequence Deletion; D014774:Virulence
chemical_list: D000935:Antifungal Agents; D016207:Cytokines; D005656:Fungal Proteins; D008364:Mannosyltransferases