Human Exonuclease 1 and BLM Helicase Interact to Resect DNA and Initiate DNA Repair

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2008 Oct 31

PMID 18971343

Citations 179

Authors

Amitabh V Nimonkar

A Zeynep Ozsoy

Jochen Genschel

Paul Modrich

Stephen C Kowalczykowski

Affiliations

Soon will be listed here.

Abstract

The error-free repair of double-stranded DNA breaks by homologous recombination requires processing of broken ends. These processed ends are substrates for assembly of DNA strand exchange proteins that mediate DNA strand invasion. Here, we establish that human BLM helicase, a member of the RecQ family, stimulates the nucleolytic activity of human exonuclease 1 (hExo1), a 5'-->3' double-stranded DNA exonuclease. The stimulation is specific because other RecQ homologs fail to stimulate hExo1. Stimulation of DNA resection by hExo1 is independent of BLM helicase activity and is, instead, mediated by an interaction between the 2 proteins. Finally, we show that DNA ends resected by hExo1 and BLM are used by human Rad51, but not its yeast or bacterial counterparts, to promote homologous DNA pairing. This in vitro system recapitulates initial steps of homologous recombination and provides biochemical evidence for a role of BLM and Exo1 in the initiation of recombinational DNA repair.

Citing Articles

HDAC4 influences the DNA damage response and counteracts senescence by assembling with HDAC1/HDAC2 to control H2BK120 acetylation and homology-directed repair.

Di Giorgio E, Dalla E, Tolotto V, DEste F, Paluvai H, Ranzino L Nucleic Acids Res. 2024; 52(14):8218-8240.

PMID: 38874468 PMC: 11317144. DOI: 10.1093/nar/gkae501.

The BLM helicase is a new therapeutic target in multiple myeloma involved in replication stress survival and drug resistance.

Ovejero S, Viziteu E, Dutrieux L, Devin J, Lin Y, Alaterre E Front Immunol. 2022; 13:983181.

PMID: 36569948 PMC: 9780552. DOI: 10.3389/fimmu.2022.983181.

Dynamic Modelling of DNA Repair Pathway at the Molecular Level: A New Perspective.

Lecca P, Ihekwaba-Ndibe A Front Mol Biosci. 2022; 9:878148.

PMID: 36177351 PMC: 9513183. DOI: 10.3389/fmolb.2022.878148.

Role of EXO1 nuclease activity in genome maintenance, the immune response and tumor suppression in Exo1D173A mice.

Wang S, Lee K, Gray S, Zhang Y, Tang C, Morrish R Nucleic Acids Res. 2022; 50(14):8093-8106.

PMID: 35849338 PMC: 9371890. DOI: 10.1093/nar/gkac616.

Targeting telomeres: advances in telomere maintenance mechanism-specific cancer therapies.

Gao J, Pickett H Nat Rev Cancer. 2022; 22(9):515-532.

PMID: 35790854 DOI: 10.1038/s41568-022-00490-1.

References

Llorente B, Symington L . The Mre11 nuclease is not required for 5' to 3' resection at multiple HO-induced double-strand breaks. Mol Cell Biol. 2004; 24(21):9682-94. PMC: 522228. DOI: 10.1128/MCB.24.21.9682-9694.2004. View

Dzantiev L, Constantin N, Genschel J, Iyer R, Burgers P, Modrich P . A defined human system that supports bidirectional mismatch-provoked excision. Mol Cell. 2004; 15(1):31-41. DOI: 10.1016/j.molcel.2004.06.016. View

Lovett S, Kolodner R . Identification and purification of a single-stranded-DNA-specific exonuclease encoded by the recJ gene of Escherichia coli. Proc Natl Acad Sci U S A. 1989; 86(8):2627-31. PMC: 286970. DOI: 10.1073/pnas.86.8.2627. View

Szankasi P, Smith G . A DNA exonuclease induced during meiosis of Schizosaccharomyces pombe. J Biol Chem. 1992; 267(5):3014-23. View

Gangloff S, Mcdonald J, Bendixen C, Arthur L, Rothstein R . The yeast type I topoisomerase Top3 interacts with Sgs1, a DNA helicase homolog: a potential eukaryotic reverse gyrase. Mol Cell Biol. 1994; 14(12):8391-8. PMC: 359378. DOI: 10.1128/mcb.14.12.8391-8398.1994. View

Mendonca V, Klepin H, Matson S . DNA helicases in recombination and repair: construction of a delta uvrD delta helD delta recQ mutant deficient in recombination and repair. J Bacteriol. 1995; 177(5):1326-35. PMC: 176740. DOI: 10.1128/jb.177.5.1326-1335.1995. View

Ellis N, Groden J, Ye T, Straughen J, Lennon D, Ciocci S . The Bloom's syndrome gene product is homologous to RecQ helicases. Cell. 1995; 83(4):655-66. DOI: 10.1016/0092-8674(95)90105-1. View

Baumann P, Benson F, West S . Human Rad51 protein promotes ATP-dependent homologous pairing and strand transfer reactions in vitro. Cell. 1996; 87(4):757-66. DOI: 10.1016/s0092-8674(00)81394-x. View

Fiorentini P, Huang K, Tishkoff D, Kolodner R, Symington L . Exonuclease I of Saccharomyces cerevisiae functions in mitotic recombination in vivo and in vitro. Mol Cell Biol. 1997; 17(5):2764-73. PMC: 232127. DOI: 10.1128/MCB.17.5.2764. View

10.

Wold M . Replication protein A: a heterotrimeric, single-stranded DNA-binding protein required for eukaryotic DNA metabolism. Annu Rev Biochem. 1997; 66:61-92. DOI: 10.1146/annurev.biochem.66.1.61. View

11.

Karow J, Chakraverty R, Hickson I . The Bloom's syndrome gene product is a 3'-5' DNA helicase. J Biol Chem. 1998; 272(49):30611-4. DOI: 10.1074/jbc.272.49.30611. View

12.

Harmon F, Kowalczykowski S . RecQ helicase, in concert with RecA and SSB proteins, initiates and disrupts DNA recombination. Genes Dev. 1998; 12(8):1134-44. PMC: 316708. DOI: 10.1101/gad.12.8.1134. View

13.

Paull T, Gellert M . The 3' to 5' exonuclease activity of Mre 11 facilitates repair of DNA double-strand breaks. Mol Cell. 1998; 1(7):969-79. DOI: 10.1016/s1097-2765(00)80097-0. View

14.

Schmutte C, Marinescu R, Sadoff M, Guerrette S, Overhauser J, Fishel R . Human exonuclease I interacts with the mismatch repair protein hMSH2. Cancer Res. 1998; 58(20):4537-42. View

15.

Tishkoff D, Amin N, Viars C, Arden K, Kolodner R . Identification of a human gene encoding a homologue of Saccharomyces cerevisiae EXO1, an exonuclease implicated in mismatch repair and recombination. Cancer Res. 1998; 58(22):5027-31. View

16.

Harmon F, DiGate R, Kowalczykowski S . RecQ helicase and topoisomerase III comprise a novel DNA strand passage function: a conserved mechanism for control of DNA recombination. Mol Cell. 1999; 3(5):611-20. DOI: 10.1016/s1097-2765(00)80354-8. View

17.

Cotta-Ramusino C, Fachinetti D, Lucca C, Doksani Y, Lopes M, Sogo J . Exo1 processes stalled replication forks and counteracts fork reversal in checkpoint-defective cells. Mol Cell. 2005; 17(1):153-9. DOI: 10.1016/j.molcel.2004.11.032. View

18.

Macris M, Krejci L, Bussen W, Shimamoto A, Sung P . Biochemical characterization of the RECQ4 protein, mutated in Rothmund-Thomson syndrome. DNA Repair (Amst). 2005; 5(2):172-80. DOI: 10.1016/j.dnarep.2005.09.005. View

19.

Wu L, Bachrati C, Ou J, Xu C, Yin J, Chang M . BLAP75/RMI1 promotes the BLM-dependent dissolution of homologous recombination intermediates. Proc Natl Acad Sci U S A. 2006; 103(11):4068-73. PMC: 1449647. DOI: 10.1073/pnas.0508295103. View

20.

Bachrati C, Borts R, Hickson I . Mobile D-loops are a preferred substrate for the Bloom's syndrome helicase. Nucleic Acids Res. 2006; 34(8):2269-79. PMC: 1456333. DOI: 10.1093/nar/gkl258. View