The Checkpoint Clamp Protein Rad9 Facilitates DNA-end Resection and Prevents Alternative Non-homologous End Joining

Overview

Journal Cell Cycle

Specialty Cell Biology

Date 2014 Dec 9

PMID 25485590

Citations 11

Authors

Feng-Ling Tsai

Mihoko Kai

Affiliations

Soon will be listed here.

Abstract

DNA damage activates the cell cycle checkpoint to regulate cell cycle progression. The checkpoint clamp (Rad9-Hus1-Rad1 complex) is recruited to damage sites, and is required for checkpoint activation. While functions of the checkpoint clamp in checkpoint activation have been well studied, its functions in DNA repair regulation remain elusive. Here we show that Rad9 is required for efficient homologous recombination (HR), and facilitates DNA-end resection. The role of Rad9 in homologous recombination is independent of its function in checkpoint activation, and this function is important for preventing alternative non-homologous end joining (altNHEJ). These findings reveal novel function of the checkpoint clamp in HR.

Citing Articles

Resection of DNA double-strand breaks activates Mre11-Rad50-Nbs1- and Rad9-Hus1-Rad1-dependent mechanisms that redundantly promote ATR checkpoint activation and end processing in Xenopus egg extracts.

Tatsukawa K, Sakamoto R, Kawasoe Y, Kubota Y, Tsurimoto T, Takahashi T Nucleic Acids Res. 2024; 52(6):3146-3163.

PMID: 38349040 PMC: 11014350. DOI: 10.1093/nar/gkae082.

DNA binding by the Rad9A subunit of the Rad9-Rad1-Hus1 complex.

Hwang B, Gonzales R, Corzine S, Stenson E, Pidugu L, Lu A PLoS One. 2022; 17(8):e0272645.

PMID: 35939452 PMC: 9359528. DOI: 10.1371/journal.pone.0272645.

Hypermethylation of intron 2 in childhood cancer patients, leukemia and tumor cell lines suggest a role for oncogenic transformation.

Galetzka D, Bock J, Wagner L, Dittrich M, Sinizyn O, Ludwig M EXCLI J. 2022; 21:117-143.

PMID: 35221838 PMC: 8859646. DOI: 10.17179/excli2021-4482.

The Dark Side of UV-Induced DNA Lesion Repair.

Strzalka W, Zglobicki P, Kowalska E, Bazant A, Dziga D, Banas A Genes (Basel). 2020; 11(12).

PMID: 33276692 PMC: 7761550. DOI: 10.3390/genes11121450.

DNMT1 and DNMT3B regulate tumorigenicity of human prostate cancer cells by controlling RAD9 expression through targeted methylation.

Zhu A, Hopkins K, Friedman R, Bernstock J, Broustas C, Lieberman H Carcinogenesis. 2020; 42(2):220-231.

PMID: 32780107 PMC: 7905840. DOI: 10.1093/carcin/bgaa088.

References

Kass E, Helgadottir H, Chen C, Barbera M, Wang R, Westermark U . Double-strand break repair by homologous recombination in primary mouse somatic cells requires BRCA1 but not the ATM kinase. Proc Natl Acad Sci U S A. 2013; 110(14):5564-9. PMC: 3619303. DOI: 10.1073/pnas.1216824110. View

Lin J, Dutta A . ATR pathway is the primary pathway for activating G2/M checkpoint induction after re-replication. J Biol Chem. 2007; 282(42):30357-62. DOI: 10.1074/jbc.M705178200. View

Gunn A, Stark J . I-SceI-based assays to examine distinct repair outcomes of mammalian chromosomal double strand breaks. Methods Mol Biol. 2012; 920:379-91. DOI: 10.1007/978-1-61779-998-3_27. View

Nimonkar A, Genschel J, Kinoshita E, Polaczek P, Campbell J, Wyman C . BLM-DNA2-RPA-MRN and EXO1-BLM-RPA-MRN constitute two DNA end resection machineries for human DNA break repair. Genes Dev. 2011; 25(4):350-62. PMC: 3042158. DOI: 10.1101/gad.2003811. View

Ellison V, Stillman B . Biochemical characterization of DNA damage checkpoint complexes: clamp loader and clamp complexes with specificity for 5' recessed DNA. PLoS Biol. 2003; 1(2):E33. PMC: 261875. DOI: 10.1371/journal.pbio.0000033. View

Shiotani B, Nguyen H, Hakansson P, Marechal A, Tse A, Tahara H . Two distinct modes of ATR activation orchestrated by Rad17 and Nbs1. Cell Rep. 2013; 3(5):1651-62. PMC: 3680100. DOI: 10.1016/j.celrep.2013.04.018. View

Chapman J, Barral P, Vannier J, Borel V, Steger M, Tomas-Loba A . RIF1 is essential for 53BP1-dependent nonhomologous end joining and suppression of DNA double-strand break resection. Mol Cell. 2013; 49(5):858-71. PMC: 3594748. DOI: 10.1016/j.molcel.2013.01.002. View

Sartori A, Lukas C, Coates J, Mistrik M, Fu S, Bartek J . Human CtIP promotes DNA end resection. Nature. 2007; 450(7169):509-14. PMC: 2409435. DOI: 10.1038/nature06337. View

Lee J, Paull T . Activation and regulation of ATM kinase activity in response to DNA double-strand breaks. Oncogene. 2007; 26(56):7741-8. DOI: 10.1038/sj.onc.1210872. View

10.

Tang J, Cho N, Cui G, Manion E, Shanbhag N, Botuyan M . Acetylation limits 53BP1 association with damaged chromatin to promote homologous recombination. Nat Struct Mol Biol. 2013; 20(3):317-25. PMC: 3594358. DOI: 10.1038/nsmb.2499. View

11.

Roos-Mattjus P, Hopkins K, Oestreich A, Vroman B, Johnson K, Naylor S . Phosphorylation of human Rad9 is required for genotoxin-activated checkpoint signaling. J Biol Chem. 2003; 278(27):24428-37. DOI: 10.1074/jbc.M301544200. View

12.

Blaikley E, Tinline-Purvis H, Kasparek T, Marguerat S, Sarkar S, Hulme L . The DNA damage checkpoint pathway promotes extensive resection and nucleotide synthesis to facilitate homologous recombination repair and genome stability in fission yeast. Nucleic Acids Res. 2014; 42(9):5644-56. PMC: 4027169. DOI: 10.1093/nar/gku190. View

13.

Liu S, Opiyo S, Manthey K, Glanzer J, Ashley A, Amerin C . Distinct roles for DNA-PK, ATM and ATR in RPA phosphorylation and checkpoint activation in response to replication stress. Nucleic Acids Res. 2012; 40(21):10780-94. PMC: 3510507. DOI: 10.1093/nar/gks849. View

14.

Shin M, Yuan M, Zhang H, Margolick J, Kai M . ATM-dependent phosphorylation of the checkpoint clamp regulates repair pathways and maintains genomic stability. Cell Cycle. 2012; 11(9):1796-803. PMC: 3372382. DOI: 10.4161/cc.20161. View

15.

Anantha R, Vassin V, Borowiec J . Sequential and synergistic modification of human RPA stimulates chromosomal DNA repair. J Biol Chem. 2007; 282(49):35910-23. DOI: 10.1074/jbc.M704645200. View

16.

Kai M . Role of the checkpoint clamp in DNA damage response. Biomolecules. 2014; 3(1):75-84. PMC: 4030880. DOI: 10.3390/biom3010075. View

17.

Ngo G, Balakrishnan L, Dubarry M, Campbell J, Lydall D . The 9-1-1 checkpoint clamp stimulates DNA resection by Dna2-Sgs1 and Exo1. Nucleic Acids Res. 2014; 42(16):10516-28. PMC: 4176354. DOI: 10.1093/nar/gku746. View

18.

Costelloe T, Louge R, Tomimatsu N, Mukherjee B, Martini E, Khadaroo B . The yeast Fun30 and human SMARCAD1 chromatin remodellers promote DNA end resection. Nature. 2012; 489(7417):581-4. PMC: 3493121. DOI: 10.1038/nature11353. View

19.

Shanbhag N, Rafalska-Metcalf I, Balane-Bolivar C, Janicki S, Greenberg R . ATM-dependent chromatin changes silence transcription in cis to DNA double-strand breaks. Cell. 2010; 141(6):970-81. PMC: 2920610. DOI: 10.1016/j.cell.2010.04.038. View

20.

Lee-Theilen M, Matthews A, Kelly D, Zheng S, Chaudhuri J . CtIP promotes microhomology-mediated alternative end joining during class-switch recombination. Nat Struct Mol Biol. 2010; 18(1):75-9. PMC: 3471154. DOI: 10.1038/nsmb.1942. View