Abstract
In response to replication block or DNA damage in S phase the DNA replication and DNA damage checkpoints are activated. The current model in human predicts, that a Rad17/Replication factor C (RF-C) complex might serve as a recruitment complex for the Rad9/Hus1/Rad1 complex to sites of replication block or DNA damage. In this study we have investigated the fate of the Rad17/RF-C complex after treatment of synchronized Hela cells with the replication inhibitor hydroxyurea. In hydroxyurea treated cells the RF-C p37 subunit became more resistant to extraction. Moreover, co-immunoprecipitation studies with extracts of hydroxyurea treated cells showed an interaction of RF-C p37 with Rad17 and of PCNA with Rad9 and RF-C p37. An enhanced colocalization of Rad17 and PCNA in late S phase after hydroxyurea treatment was observed. Our data suggested, that upon replication block a Rad17/RF-C complex is recruited to sites of DNA lesions in late S phase, binds the Rad9/Hus1/Rad1 complex and enables it to interact with PCNA. An interaction of Rad17/RF-C with PCNA appears to be mediated by the small RF-C p37 subunit, suggesting that PCNA might provide communication between replication checkpoint control and DNA replication and repair.
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References
Bao S, Tibbetts RS, Brumbaugh KM, Fang Y, Richardson DA, Ali A, Chen SM, Abraham RT, Wang XF . 2001 Nature 411: 969–974
Bessho T, Sancar A . 2000 J. Biol. Chem. 275: 7451–7454
Bravo R, MacDonald-Bravo H . 1985 EMBO J. 4: 655–661
Bravo R . 1986 Exp. Cell Res. 163: 287–293
Bravo R, MacDonald-Bravo H . 1987 J. Cell Biol. 105: 1549–1554
Burtelow M, Roos-Mattjus P, Rauen M, Babendure J, Karnitz L . 2001 J. Biol. Chem. 276: 25903–25909
Chang MS, Sasaki H, Campbell MS, Kraeft SK, Sutherland R, Yang CY, Liu Y, Auclair D, Hao L, Sonoda H, Ferland LH, Chen LB . 1999 J. Biol. Chem. 274: 36544–36549
Chen MS, Higashikubo R, Laszlo A, Roti Roti J . 2001a Gene 277: 145–152
Chen MJ, Lin YT, Lieberman HB, Chen G, Lee EY . 2001b J. Biol. Chem. 276: 16580–16586
de la Torre-Ruiz MA, Green CM, Lowndes NF . 1998 EMBO J. 17: 2687–2698
Donaldson AD, Blow JJ . 2001 Curr. Biol. 11: R979–R982
Fox MH, Arndt-Jovin DJ, Jovin TM, Baumann PH, Robert-Nicoud M . 1991 J. Cell Sci. 99: 247–253
Gary Schmidt SL, Pautz AL, Burgers PM . 2001 J. Biol. Chem. 276: 34792–34800
Green CM, Erdjument-Bromage H, Tempst P, Lowndes NF . 2000 Curr. Biol. 10: 39–42
Griffith JD, Lindsey-Boltz LA, Sancar A . 2002 J. Biol. Chem. 277: 15233–15236
Kai M, Tanaka H, Wang TS . 2001 Mol. Cell. Biol. 21: 3289–3301
Komatsu K, Wharton W, Hang H, Wu C, Singh S, Lieberman HB, Pledger WJ, Wang HG . 2000 Oncogene 19: 5291–5297
Lindsay HD, Griffiths DJ, Edwards RJ, Christensen PU, Murray JM, Osman F, Walworth N, Carr AM . 1998 Genes Dev. 12: 382–395
Lindsey-Boltz LA, Bermudez VP, Hurwitz J, Sancar A . 2001 Proc. Natl. Acad. Sci. USA 98: 11236–11241
Loor G, Zhang SJ, Zhang P, Toomey NL, Lee MY . 1997 Nucleic Acids Res. 25: 5041–5046
Lopes M, Cotta-Ramusino C, Pellicioli A, Liberi G, Plevani P, Muzi-Falconi M, Newlon CS, Foiani M . 2001 Nature 412: 557–561
Mitchell A . 2001 Nat. Rev. Mol. Cell Biol. 2: 563
Miura M . 1999 J. Radiat. Res. (Tokyo) 40: 1–12
Mossi R, Jonsson ZO, Allen BL, Hardin SH, Hübscher U . 1997 J. Biol. Chem. 272: 1769–1776
Mossi R, Hübscher U . 1998 Eur. J. Biochem. 254: 209–216
Naiki T, Shimomura T, Kondo T, Matsumoto K, Sugimoto K . 2000 Mol. Cell. Biol. 20: 5888–5896
Noskov VN, Araki H, Sugino A . 1998 Mol. Cell. Biol. 18: 4914–4923
Parker AE, Van de Weyer I, Laus MC, Oostveen I, Yon J, Verhasselt P, Luyten WH . 1998 J. Biol. Chem. 273: 18332–18339
Rauen M, Burtelow MA, Dufault VM, Karnitz LM . 2000 J. Biol. Chem. 275: 29767–29771
Reynolds N, Fantes PA, MacNeill SA . 1999 Nucleic Acids Res. 27: 462–469
Santocanale C, Diffley JF . 1998 Nature 395: 615–618
Sasaki K, Kurose A, Ishida Y . 1993 Cytometry 14: 876–882
Shimada M, Okuzaki D, Tanaka S, Tougan T, Tamai KK, Shimoda C, Nojima H . 1999 Mol. Biol. Cell 10: 3991–4003
Shimomura T, Ando S, Matsumoto K, Sugimoto K . 1998 Mol. Cell. Biol. 18: 5485–5491
Shirahige K, Hori Y, Shiraishi K, Yamashita M, Takahashi K, Obuse C, Tsurimoto T, Yoshikawa H . 1998 Nature 395: 618–621
Sugimoto K, Shimomura T, Hashimoto K, Araki H, Sugino A, Matsumoto K . 1996 Proc. Natl. Acad. Sci. USA 93: 7048–7052
Sugimoto K, Ando S, Shimomura T, Matsumoto K . 1997 Mol. Cell. Biol. 17: 5905–5914
Tercero JA, Diffley JF . 2001 Nature 412: 553–557
Tsurimoto T . 1999 Front. Biosci. 4: D849–D858
Venclovas C, Thelen MP . 2000 Nucleic Acids Res. 28: 2481–2493
Walworth NC, Bernards R . 1996 Science 271: 353–356
Zou L, Cortez D, Elledge SJ . 2002 Genes Dev. 16: 198–208
Acknowledgements
We thank S Keller for technical support and advice, S Koundrioukoff for help with cell culture and T Bächi, AG Bittermann and M Höchli for their support with confocal microscopy. We are indebted to M Foiani for his critical reading of the manuscript. This study was supported by the Swiss National Science Foundation (grant 3100. 061361. 00), by Cancer League of the Kanton Zürich and by the Kanton of Zürich.
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Dahm, K., Hübscher, U. Colocalization of human Rad17 and PCNA in late S phase of the cell cycle upon replication block. Oncogene 21, 7710–7719 (2002). https://doi.org/10.1038/sj.onc.1205872
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DOI: https://doi.org/10.1038/sj.onc.1205872