Purification and characterization of human DNA damage checkpoint Rad complexes Journal Article

   


Authors: Lindsey-Boltz, L. A.; Bermudez, V. P.; Hurwitz, J.; Sancar, A.
Article Title: Purification and characterization of human DNA damage checkpoint Rad complexes
Abstract: Checkpoint Rad proteins function early in the DNA damage checkpoint signaling cascade to arrest cell cycle progression in response to DNA damage. This checkpoint ensures the transmission of an intact genetic complement to daughter cells. To learn about the damage sensor function of the human checkpoint Rad proteins, we purified a heteropentameric complex composed of hRad17-RFCp36-RFCp37-RFCp38-RFCp40 (hRad17-RFC) and a heterotrimeric complex composed of hRad9-hHus1-hRad1 (checkpoint 9-1-1 complex). hRad17RFC binds to DNA, with a preference for primed DNA and possesses weak ATPase activity that is stimulated by primed DNA and singlestranded DNA. hRad17-RFC forms a complex with the 9-1-1 heterotrimer reminiscent of the replication factor C/proliferating cell nuclear antigen clamp loader/sliding clamp complex of the replication machinery. These findings constitute biochemical support for models regarding the roles of checkpoint Rads as damage sensors in the DNA damage checkpoint response of human cells.
Keywords: signal transduction; controlled study; unclassified drug; human cell; dna-binding proteins; nonhuman; dna replication; animal cell; cell cycle proteins; dna damage; cell cycle; complex formation; embryo; protein dna binding; cell line; transcription, genetic; hela cells; transfection; animalia; dna; kinetics; enzyme analysis; recombinant proteins; protein biosynthesis; binding sites; replication factor c; cycline; single stranded dna; protein subunits; replication protein c; adenosine triphosphatase; protein kinase; adenosine triphosphatases; enzyme purification; exonucleases; macromolecular substances; genetic complementation; cell-free system; humans; human; priority journal; article; protein rad
Journal Title: Proceedings of the National Academy of Sciences of the United States of America
Volume: 98
Issue: 20
ISSN: 0027-8424
Publisher: National Academy of Sciences  
Date Published: 2001-09-25
Start Page: 11236
End Page: 11241
Language: English
DOI: 10.1073/pnas.201373498
PUBMED: 11572977
PROVIDER: scopus
PMCID: PMC58713
DOI/URL:

http://www.scopus.com/inward/record.url?eid=2-s2.0-0035949503&partnerID=40&md5=296c787bd305e017d00c7f3c6561334a
Notes: Export Date: 21 May 2015 -- Source: Scopus
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MSK Authors
  1. Vladimir D Bermudez
    32 Bermudez
  2. Jerard Hurwitz
    206 Hurwitz
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