title: CK2 phosphorylates SSRP1 and inhibits its DNA-binding activity.
abstract: We have previously shown that CK2 associates with the human high-mobility group protein SSRP1 and that this association increases in response to UV irradiation. CK2 also phosphorylates SSRP1 in vitro. Here we extend this work by investigating CK2 regulation of SSRP1 function through phosphorylation. Phosphorylation of SSRP1 by CK2 inhibited the nonspecific DNA-binding activity of SSRP1 and FACT (facilitating chromatin-mediated transcription) complex in vitro. Using a serine/threonine-scanning Auto-spot peptide array coupled with a filter-based kinase assay with synthetic peptides as substrates, we identified serines 510, 657, and 688 as phosphorylation targets of CK2 in vitro. Mutagenesis of the three serines revealed that serine 510 was more important for the regulation of SSRP1 DNA-binding activity. Furthermore, we found that SSRP1 was phosphorylated in cells in response to UV (but not gamma) irradiation. These results suggest that CK2 regulates the DNA-binding ability of SSRP1 and that this regulation may be responsive to specific cell stresses.
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Portland, Oregon 97239, USA.
authors.forename: Yanping
authors.initials: Y
authors.lastname: Li
authors.forename: David M
authors.initials: DM
authors.lastname: Keller
authors.forename: John D
authors.initials: JD
authors.lastname: Scott
authors.forename: Hua
authors.initials: H
authors.lastname: Lu
mesh_terms: D047390:Casein Kinase II / Q000502:physiology*; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006367:HeLa Cells; D006609:High Mobility Group Proteins / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D012694:Serine / Q000378:metabolism; D035602:Transcriptional Elongation Factors / Q000378:metabolism*; D014466:Ultraviolet Rays
chemical_list: D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; C073537:SSRP1 protein, human; D035602:Transcriptional Elongation Factors; D012694:Serine; D004247:DNA; D047390:Casein Kinase II