Mapping of amino acid residues in the p34 subunit of human single- stranded DNA-binding protein phosphorylated by DNA-dependent protein kinase and Cdc2 kinase in vitro Journal Article

  


Authors: Niu, H.; Erdjument-Bromage, H.; Pan, Z. Q.; Lee, S. H.; Tempst, P.; Hurwitz, J.
Article Title: Mapping of amino acid residues in the p34 subunit of human single- stranded DNA-binding protein phosphorylated by DNA-dependent protein kinase and Cdc2 kinase in vitro
Abstract: Human single-stranded DNA-binding protein (HSSB, also called RPA), is a heterotrimeric complex that consists of three subunits, p70, p34, and p11. HSSB is essential for the in vitro replication of SV40 DNA and nucleotide excision repair. It also has important functions in other DNA transactions, including DNA recombination, transcription, and double-stranded DNA break repair. The p34 subunit of HSSB is phosphorylated in a cell cycle-dependent manner. Both Cdc2 kinase and the DNA-dependent protein kinase (DNA-PK) phosphorylate HSSB-p34 in vitro. In this study, we show that efficient phosphorylation of HSSB-p34 by DNA-PK requires Ku as well as DNA. The DNA-PK phosphorylation sites in HSSB-p34 have been mapped at Thr-21 and Ser-33. Kinetic studies demonstrated that a phosphate residue is first incorporated at Thr-21 followed by the incorporation of a second phosphate residue at Ser- 33. We also identified Ser-29 as the major Cdc2 kinase phosphorylation site in the p34 subunit.
Keywords: protein phosphorylation; dna binding protein; dna-binding proteins; protein conformation; protein analysis; dna repair; hela cells; phosphorylation; transcription factors; nuclear proteins; amino acid sequence; molecular sequence data; protein-serine-threonine kinases; chromatography, high pressure liquid; single stranded dna; protein structure; structure analysis; protein kinase; dna helicases; dna-activated protein kinase; cdc2 protein kinase; antigens, nuclear; amino acid composition; humans; human; priority journal; article
Journal Title: Journal of Biological Chemistry
Volume: 272
Issue: 19
ISSN: 0021-9258
Publisher: American Society for Biochemistry and Molecular Biology  
Date Published: 1997-05-09
Start Page: 12634
End Page: 12641
Language: English
DOI: 10.1074/jbc.272.19.12634
PUBMED: 9139719
PROVIDER: scopus
DOI/URL:

https://www.scopus.com/inward/record.uri?eid=2-s2.0-0030924695&doi=10.1074%2fjbc.272.19.12634&partnerID=40&md5=0cfb2a33ecc93692a66b268543113098
Notes: Article -- Export Date: 17 March 2017 -- Source: Scopus
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MSK Authors
  1. Jerard Hurwitz
    206 Hurwitz
  2. Paul J Tempst
    324 Tempst
  3. Hediye Erdjument-Bromage
    271 Erdjument-Bromage
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