Biochemical characterization of DNA damage checkpoint complexes: Clamp loader and clamp complexes with specificity for 5' recessed DNA

Ellison, V., Stillman, B. (November 2003) Biochemical characterization of DNA damage checkpoint complexes: Clamp loader and clamp complexes with specificity for 5' recessed DNA. Plos Biology, 1 (2). pp. 231-243. ISSN 1544-9173

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DOI: 10.1371/journal.pbio.0000033


The cellular pathways involved in maintaining genome stability halt cell cycle progression in the presence of DNA damage or incomplete replication. Proteins required for this pathway include Rad17, Rad9, Hus1, Rad1, and Rfc-2, Rfc-3, Rfc-4, and Rfc-5. The heteropentamer replication factor C (RFC) loads during DNA replication the homotrimer proliferating cell nuclear antigen [PCNA) polymerase clamp onto DNA. Sequence similarities suggest the biochemical functions of an RSR (Rad17-Rfc2-Rfc3-Rfc4-Rfc5) complex and an RHR heterotrimer (Rad1-Hus1-Rad9) may be similar to that of RFC and PCNA, respectively. RSR purified from human cells loads RHR onto DNA in an ATP-, replication protein A-, and DNA structure-dependent manner. Interestingly, RSR and RFC differed in their ATPase activities and displayed distinct DNA substrate specificities. RSR preferred DNA substrates possessing 5' recessed ends whereas RFC preferred 3' recessed end DNA substrates. Characterization of the biochemical loading reaction executed by the checkpoint clamp loader RSR suggests new insights into the mechanisms underlying recognition of damage-induced DNA structures and signaling to cell cycle controls. The observation that RSR loads its clamp onto a 5' recessed end supports a potential role for RHR and RSR in diverse DNA metabolism, such as stalled DNA replication forks, recombination-linked DNA repair, and telomere maintenance, among other processes.

Item Type: Paper
Uncontrolled Keywords: REPLICATION FACTOR-C Replication Factor C POLYMERASE-III HOLOENZYME polymerase III holoenzyme CELL NUCLEAR ANTIGEN cell nuclear antigen SINGLE-STRANDED-DNA single stranded DNA FISSION YEAST RAD17 fission yease RAD17 SLIDING CLAMP sliding clamp SACCHAROMYCES-CEREVISIAE Saccharomyces Cerevisiae IN-VITRO In vitro ACCESSORY PROTEINS accessory proteins PRIMER-TEMPLATE primer template
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date: November 2003
Date Deposited: 06 Mar 2012 16:33
Last Modified: 20 Jun 2017 18:18
PMCID: PMC261875
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