title: In vivo putative O-GlcNAcylation of human SCP1 and evidence for possible role of its N-terminal disordered structure.
abstract: RNA polymerase II carboxyl-terminal domain (RNAPII CTD) phosphatases are responsible for the dephosphorylation of the C-terminal domain of the small subunit of RNAPII in eukaryotes. Recently, we demonstrated the identification of several interacting partners with human small CTD phosphatase1 (hSCP1) and the substrate specificity to delineate an appearance of the dephosphorylation catalyzed by SCP1. In this study, using the established cells for inducibly expressing hSCP1 proteins, we monitored the modification of β-O-linked N-acetylglucosamine (O-GlcNAc). O-GlcNAcylation is one of the most common post-translational modifications (PTMs). To gain insight into the PTM of hSCP1, we used the Western blot, immunoprecipitation, succinylayed wheat germ agglutininprecipitation, liquid chromatography-mass spectrometry analyses, and site-directed mutagenesis and identified the Ser41 residue of hSCP1 as the O-GlcNAc modification site. These results suggest that hSCP1 may be an O-GlcNAcylated protein in vivo, and its N-terminus may function a possible role in the PTM, providing a scaffold for binding the protein(s).
authors.affiliation: Department of Brain Science, Daegu-Gyeongbuk Institute of Science and Technology (DGIST), Daegu 711-873, Korea.
authors.forename: JaeHyung
authors.initials: J
authors.lastname: Koo
authors.affiliation: Department of Biotechnology, Konkuk University, Chungju 380-701, Korea.
authors.forename: Young Yil
authors.initials: YY
authors.lastname: Bahk
mesh_terms: D000818:Animals; D006031:Glycosylation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D064267:Intrinsically Disordered Proteins / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism*; D010749:Phosphoprotein Phosphatases / Q000737:chemistry* / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012694:Serine / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D064267:Intrinsically Disordered Proteins; D009687:Nuclear Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D012694:Serine; C413483:CTDSP1 protein, human; D010749:Phosphoprotein Phosphatases
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