gfsA encodes a novel galactofuranosyltransferase involved in biosynthesis of galactofuranose antigen of O-glycan in Aspergillus nidulans and Aspergillus fumigatus

Yuji Komachi, Shintaro Hatakeyama, Haruka Motomatsu, Taiki Futagami, Karina Kizjakina, Pablo Sobrado, Keisuke Ekino, Kaoru Takegawa, Masatoshi Goto, Yoshiyuki Nomura, Takuji Oka

Research output: Contribution to journalArticle

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Abstract

Summary: The cells walls of filamentous fungi in the genus Aspergillus have galactofuranose (Galf)-containing polysaccharides and glycoconjugates, including O-glycans, N-glycans, fungal-type galactomannan and glycosylinositolphosphoceramide, which are important for cell wall integrity. Here, we attempted to identify galactofuranosyltransferases that couple Galf monomers onto other wall components in Aspergillus nidulans. Using reverse-genetic and biochemical approaches, we identified that the AN8677 gene encoded a galactofuranosyltransferase, which we called GfsA, involved in Galf antigen biosynthesis. Disruption of gfsA reduced binding of β-Galf-specific antibody EB-A2 to O-glycosylated WscA protein and galactomannoproteins. The results of an in-vitroGalf antigen synthase assay revealed that GfsA has β1,5- or β1,6-galactofuranosyltransferase activity for O-glycans in glycoproteins, uses UDP-d-Galf as a sugar donor, and requires a divalent manganese cation for activity. GfsA was found to be localized at the Golgi apparatus based on cellular fractionation experiments. ΔgfsA cells exhibited an abnormal morphology characterized by poor hyphal extension, hyphal curvature and limited formation of conidia. Several gfsA orthologues were identified in members of the Pezizomycotina subphylum of Ascomycota, including the human pathogen Aspergillus fumigatus. To our knowledge, this is the first characterization of a fungal β-galactofuranosyltransferase, which was shown to be involved in Galf antigen biosynthesis of O-glycans in the Golgi.

Original languageEnglish
Pages (from-to)1054-1073
Number of pages20
JournalMolecular Microbiology
Volume90
Issue number5
DOIs
Publication statusPublished - Dec 1 2013

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Aspergillus nidulans
O Antigens
Aspergillus fumigatus
Polysaccharides
Cell Wall
Antigens
Reverse Genetics
Ascomycota
Glycoconjugates
Fungal Spores
Divalent Cations
Golgi Apparatus
Aspergillus
Manganese
varespladib methyl
Glycoproteins
Fungi
Antibodies
Genes
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Microbiology

Cite this

gfsA encodes a novel galactofuranosyltransferase involved in biosynthesis of galactofuranose antigen of O-glycan in Aspergillus nidulans and Aspergillus fumigatus. / Komachi, Yuji; Hatakeyama, Shintaro; Motomatsu, Haruka; Futagami, Taiki; Kizjakina, Karina; Sobrado, Pablo; Ekino, Keisuke; Takegawa, Kaoru; Goto, Masatoshi; Nomura, Yoshiyuki; Oka, Takuji.

In: Molecular Microbiology, Vol. 90, No. 5, 01.12.2013, p. 1054-1073.

Research output: Contribution to journalArticle

Komachi, Y, Hatakeyama, S, Motomatsu, H, Futagami, T, Kizjakina, K, Sobrado, P, Ekino, K, Takegawa, K, Goto, M, Nomura, Y & Oka, T 2013, 'gfsA encodes a novel galactofuranosyltransferase involved in biosynthesis of galactofuranose antigen of O-glycan in Aspergillus nidulans and Aspergillus fumigatus', Molecular Microbiology, vol. 90, no. 5, pp. 1054-1073. https://doi.org/10.1111/mmi.12416
Komachi, Yuji ; Hatakeyama, Shintaro ; Motomatsu, Haruka ; Futagami, Taiki ; Kizjakina, Karina ; Sobrado, Pablo ; Ekino, Keisuke ; Takegawa, Kaoru ; Goto, Masatoshi ; Nomura, Yoshiyuki ; Oka, Takuji. / gfsA encodes a novel galactofuranosyltransferase involved in biosynthesis of galactofuranose antigen of O-glycan in Aspergillus nidulans and Aspergillus fumigatus. In: Molecular Microbiology. 2013 ; Vol. 90, No. 5. pp. 1054-1073.
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AU - Hatakeyama, Shintaro

AU - Motomatsu, Haruka

AU - Futagami, Taiki

AU - Kizjakina, Karina

AU - Sobrado, Pablo

AU - Ekino, Keisuke

AU - Takegawa, Kaoru

AU - Goto, Masatoshi

AU - Nomura, Yoshiyuki

AU - Oka, Takuji

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