Abstract
The human Rad9-Hus1-Rad1 (9-1-1) and Rad17-RFC40-RFC38-RFC37-RFC36 (hRad17-RFC) protein complexes are important components of the mammalian deoxyribonucleic acid (DNA) damage response. Accumulating evidence from biochemical and molecular modeling studies indicate that these two complexes function as a clamp-clamp loader pair (1). The 9-1-1 complex is converted to a less-extractable, DNA-bound form after DNA damage (2). DNA damage-inducible binding of the 9-1-1 complex to chromatin is dependent on hRad17, but independent of ataxia-telangiectasia and Rad3 (ATR) (3). These data support a model in which hRad17-RFC and 9-1-1 are essential components of a DNA damage sensor that functions early in the DNA damage checkpoint pathway.
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References
O’Connell, M. J., Walworth, N. C., and Carr, A. M. (2000) The G2-phase DNA-damage checkpoint. Trends Cell Biol. 10, 296–303.
Burtelow, M. A., Kaufmann, S. H., and Karnitz, L. M. (2000) Retention of the human Rad9 checkpoint complex in extraction-resistant nuclear complexes after DNA damage. J. Biol. Chem. 275, 26,343–26,348.
Zou, L., Cortez, D., and Elledge, S. J. (2002) Regulation of ATR substrate selection by Rad17-dependent loading of Rad9 complexes onto chromatin Genes Dev. 16, 198–208.
Podust, V. N., Tiwari, N., Stephan, S., and Fanning, E. (1998) Replication factor C disengages from proliferating cell nuclear antigen (PCNA) upon sliding clamp formation, and PCNA itself tethers DNA polymerase delta to DNA. J. Biol. Chem. 273, 31,992–31,999.
Podust, V. N., and Fanning, E. (1997) Assembly of functional replication factor C expressed using recombinant baculoviruses. J. Biol. Chem. 272, 6303–6310.
Burtelow, M. A., Roos-Mattjus, P. M., Rauen, M., Babendure, J. R., and Karnitz, L. M. (2001) Reconstitution and molecular analysis of the hRad9-hHus1-hRad1 (9-1-1) DNA damage responsive checkpoint complex. J. Biol. Chem. 276, 25,903–25,909.
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© 2004 Humana Press Inc., Totowa, NJ
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Burtelow, M.A., Podust, V.N., Karnitz, L.M. (2004). Purification and Identification of Protein Complexes That Control the Cell Cycle. In: Lieberman, H.B. (eds) Cell Cycle Checkpoint Control Protocols. Methods in Molecular Biology™, vol 241. Humana Press. https://doi.org/10.1385/1-59259-646-0:247
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DOI: https://doi.org/10.1385/1-59259-646-0:247
Publisher Name: Humana Press
Print ISBN: 978-1-58829-115-8
Online ISBN: 978-1-59259-646-1
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