title: Discovery of a nucleocytoplasmic O-mannose glycoproteome in yeast.
abstract: Dynamic cycling of N-Acetylglucosamine (GlcNAc) on serine and threonine residues (O-GlcNAcylation) is an essential process in all eukaryotic cells except yeast, including Saccharomyces cerevisiae and Schizosaccharomyces pombe. O-GlcNAcylation modulates signaling and cellular processes in an intricate interplay with protein phosphorylation and serves as a key sensor of nutrients by linking the hexosamine biosynthetic pathway to cellular signaling. A longstanding conundrum has been how yeast survives without O-GlcNAcylation in light of its similar phosphorylation signaling system. We previously developed a sensitive lectin enrichment and mass spectrometry workflow for identification of the human O-linked mannose (O-Man) glycoproteome and used this to identify a pleothora of O-Man glycoproteins in human cell lines including the large family of cadherins and protocadherins. Here, we applied the workflow to yeast with the aim to characterize the yeast O-Man glycoproteome, and in doing so, we discovered hitherto unknown O-Man glycosites on nuclear, cytoplasmic, and mitochondrial proteins in S. cerevisiae and S. pombe. Such O-Man glycoproteins were not found in our analysis of human cell lines. However, the type of yeast O-Man nucleocytoplasmic proteins and the localization of identified O-Man residues mirror that of the O-GlcNAc glycoproteome found in other eukaryotic cells, indicating that the two different types of O-glycosylations serve the same important biological functions. The discovery opens for exploration of the enzymatic machinery that is predicted to regulate the nucleocytoplasmic O-Man glycosylations. It is likely that manipulation of this type of O-Man glycosylation will have wide applications for yeast bioprocessing.
authors.affiliation: Copenhagen Center for Glycomics, Departments of Cellular and Molecular Medicine, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark; hclau@sund.ku.dk halim@sund.ku.dk.
authors.forename: Adnan
authors.initials: A
authors.lastname: Halim
authors.affiliation: Copenhagen Center for Glycomics, Departments of Cellular and Molecular Medicine, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark;
authors.forename: Ida Signe Bohse
authors.initials: IS
authors.lastname: Larsen
authors.affiliation: Centre for Organismal Studies (COS), University of Heidelberg, D-69120 Heidelberg, Germany;
authors.forename: Patrick
authors.initials: P
authors.lastname: Neubert
authors.affiliation: Copenhagen Center for Glycomics, Departments of Cellular and Molecular Medicine, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark;
authors.forename: Hiren Jitendra
authors.initials: HJ
authors.lastname: Joshi
authors.affiliation: Copenhagen Center for Glycomics, Departments of Cellular and Molecular Medicine, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark; Department of Plant and Environmental Sciences, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.
authors.forename: Bent Larsen
authors.initials: BL
authors.lastname: Petersen
authors.affiliation: Copenhagen Center for Glycomics, Departments of Cellular and Molecular Medicine, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark;
authors.forename: Sergey Y
authors.initials: SY
authors.lastname: Vakhrushev
authors.affiliation: Centre for Organismal Studies (COS), University of Heidelberg, D-69120 Heidelberg, Germany;
authors.forename: Sabine
authors.initials: S
authors.lastname: Strahl
authors.affiliation: Copenhagen Center for Glycomics, Departments of Cellular and Molecular Medicine, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark; hclau@sund.ku.dk halim@sund.ku.dk.
authors.forename: Henrik
authors.initials: H
authors.lastname: Clausen
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D008358:Mannose / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D020543:Proteome; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000378:metabolism*
chemical_list: D006023:Glycoproteins; D020543:Proteome; D008358:Mannose; D000117:Acetylglucosamine
keywords: O-glycosylation; glycoproteomics; mass spectrometry; signaling; yeast