Regulation of ATR Substrate Selection by Rad17-dependent Loading of Rad9 Complexes Onto Chromatin

Overview

Journal Genes Dev

Specialty Molecular Biology

Date 2002 Jan 19

PMID 11799063

Citations 212

Authors

Lee Zou

David Cortez

Stephen J Elledge

Affiliations

Soon will be listed here.

Abstract

Cells respond to DNA damage by activating a network of signaling pathways that control cell cycle progression and DNA repair. Genetic studies in yeast suggested that several checkpoint proteins, including the RFC-related Rad17 protein, and the PCNA-related Rad1-Rad9-Hus1 protein complex might function as sensors of DNA damage. In this study, we show that the human Rad17 protein recruits the Rad9 protein complex onto chromatin after damage. Rad17 binds to chromatin prior to damage and is phosphorylated by ATR on chromatin after damage but Rad17's phosphorylation is not required for Rad9 loading onto chromatin. The chromatin associations of Rad17 and ATR are largely independent, which suggests that they localize to DNA damage independently. Furthermore, the phosphorylation of Rad17 requires Hus1, suggesting that the Rad1-Rad9-Hus1 complex recruited by Rad17 enables ATR to recognize its substrates. Our data are consistent with a model in which multiple checkpoint protein complexes localize to sites of DNA damage independently and interact to trigger the checkpoint-signaling cascade.

Citing Articles

Separable roles of the DNA damage response kinase Mec1ATR and its activator Rad24RAD17 during meiotic recombination.

Crawford M, Harper J, Cooper T, Marsolier-Kergoat M, Llorente B, Neale M PLoS Genet. 2024; 20(12):e1011485.

PMID: 39652586 PMC: 11658708. DOI: 10.1371/journal.pgen.1011485.

The Potential for Targeting G/M Cell Cycle Checkpoint Kinases in Enhancing the Efficacy of Radiotherapy.

Melia E, Parsons J Cancers (Basel). 2024; 16(17).

PMID: 39272874 PMC: 11394570. DOI: 10.3390/cancers16173016.

The abscission checkpoint senses chromatin bridges through Top2α recruitment to DNA knots.

Petsalaki E, Balafouti S, Kyriazi A, Zachos G J Cell Biol. 2023; 222(11).

PMID: 37638884 PMC: 10461104. DOI: 10.1083/jcb.202303123.

Therapeutic Targeting of DNA Replication Stress in Cancer.

Gu L, Hickey R, Malkas L Genes (Basel). 2023; 14(7).

PMID: 37510250 PMC: 10378776. DOI: 10.3390/genes14071346.

Rad17 Translocates to Nucleolus upon UV Irradiation through Nucleolar Localization Signal in the Central Basic Domain.

Fukumoto Y, Ikeuchi M, Nakayama Y, Ogra Y Int J Mol Sci. 2022; 23(20).

PMID: 36293155 PMC: 9603387. DOI: 10.3390/ijms232012300.

References

Tibbetts R, Cortez D, Brumbaugh K, Scully R, Livingston D, Elledge S . Functional interactions between BRCA1 and the checkpoint kinase ATR during genotoxic stress. Genes Dev. 2000; 14(23):2989-3002. PMC: 317107. DOI: 10.1101/gad.851000. View

Griffiths D, Barbet N, Mccready S, Lehmann A, Carr A . Fission yeast rad17: a homologue of budding yeast RAD24 that shares regions of sequence similarity with DNA polymerase accessory proteins. EMBO J. 1995; 14(23):5812-23. PMC: 394699. DOI: 10.1002/j.1460-2075.1995.tb00269.x. View

Edwards R, Bentley N, Carr A . A Rad3-Rad26 complex responds to DNA damage independently of other checkpoint proteins. Nat Cell Biol. 1999; 1(7):393-8. DOI: 10.1038/15623. View

Elbashir S, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T . Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 2001; 411(6836):494-8. DOI: 10.1038/35078107. View

Zhao H, Piwnica-Worms H . ATR-mediated checkpoint pathways regulate phosphorylation and activation of human Chk1. Mol Cell Biol. 2001; 21(13):4129-39. PMC: 87074. DOI: 10.1128/MCB.21.13.4129-4139.2001. View

Bao S, Tibbetts R, Brumbaugh K, Fang Y, Richardson D, Ali A . ATR/ATM-mediated phosphorylation of human Rad17 is required for genotoxic stress responses. Nature. 2001; 411(6840):969-74. DOI: 10.1038/35082110. View

Burtelow M, Rauen M, Babendure J, Karnitz L . Reconstitution and molecular analysis of the hRad9-hHus1-hRad1 (9-1-1) DNA damage responsive checkpoint complex. J Biol Chem. 2001; 276(28):25903-9. DOI: 10.1074/jbc.M102946200. View

Makiniemi M, Hillukkala T, Tuusa J, Reini K, Vaara M, Huang D . BRCT domain-containing protein TopBP1 functions in DNA replication and damage response. J Biol Chem. 2001; 276(32):30399-406. DOI: 10.1074/jbc.M102245200. View

Bermudez V, Hurwitz J, Sancar A . Purification and characterization of human DNA damage checkpoint Rad complexes. Proc Natl Acad Sci U S A. 2001; 98(20):11236-41. PMC: 58713. DOI: 10.1073/pnas.201373498. View

10.

Kondo T, Wakayama T, Naiki T, Matsumoto K, Sugimoto K . Recruitment of Mec1 and Ddc1 checkpoint proteins to double-strand breaks through distinct mechanisms. Science. 2001; 294(5543):867-70. DOI: 10.1126/science.1063827. View

11.

Melo J, Cohen J, Toczyski D . Two checkpoint complexes are independently recruited to sites of DNA damage in vivo. Genes Dev. 2001; 15(21):2809-21. PMC: 312815. DOI: 10.1101/gad.903501. View

12.

Alcasabas A, Osborn A, Bachant J, Hu F, Werler P, Bousset K . Mrc1 transduces signals of DNA replication stress to activate Rad53. Nat Cell Biol. 2001; 3(11):958-65. DOI: 10.1038/ncb1101-958. View

13.

Cortez D, Guntuku S, Qin J, Elledge S . ATR and ATRIP: partners in checkpoint signaling. Science. 2001; 294(5547):1713-6. DOI: 10.1126/science.1065521. View

14.

Nelms B, Maser R, Mackay J, Lagally M, Petrini J . In situ visualization of DNA double-strand break repair in human fibroblasts. Science. 1998; 280(5363):590-2. DOI: 10.1126/science.280.5363.590. View

15.

Parker A, Van de Weyer I, Laus M, Oostveen I, Yon J, Verhasselt P . A human homologue of the Schizosaccharomyces pombe rad1+ checkpoint gene encodes an exonuclease. J Biol Chem. 1998; 273(29):18332-9. DOI: 10.1074/jbc.273.29.18332. View

16.

Parker A, Van de Weyer I, Laus M, Verhasselt P, Luyten W . Identification of a human homologue of the Schizosaccharomyces pombe rad17+ checkpoint gene. J Biol Chem. 1998; 273(29):18340-6. DOI: 10.1074/jbc.273.29.18340. View

17.

Cai J, Yao N, Gibbs E, Finkelstein J, Phillips B, ODonnell M . ATP hydrolysis catalyzed by human replication factor C requires participation of multiple subunits. Proc Natl Acad Sci U S A. 1998; 95(20):11607-12. PMC: 21688. DOI: 10.1073/pnas.95.20.11607. View

18.

Liu Q, Li M, Leibham D, Cortez D, Elledge S . The univector plasmid-fusion system, a method for rapid construction of recombinant DNA without restriction enzymes. Curr Biol. 1998; 8(24):1300-9. DOI: 10.1016/s0960-9822(07)00560-x. View

19.

Volkmer E, Karnitz L . Human homologs of Schizosaccharomyces pombe rad1, hus1, and rad9 form a DNA damage-responsive protein complex. J Biol Chem. 1999; 274(2):567-70. DOI: 10.1074/jbc.274.2.567. View

20.

Thelen M, Venclovas C, Fidelis K . A sliding clamp model for the Rad1 family of cell cycle checkpoint proteins. Cell. 1999; 96(6):769-70. DOI: 10.1016/s0092-8674(00)80587-5. View