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BLM-DNA2-RPA-MRN and EXO1-BLM-RPA-MRN constitute two DNA end resection machineries for human DNA break repair.

Publication ,  Journal Article
Nimonkar, AV; Genschel, J; Kinoshita, E; Polaczek, P; Campbell, JL; Wyman, C; Modrich, P; Kowalczykowski, SC
Published in: Genes Dev
February 15, 2011

Repair of dsDNA breaks requires processing to produce 3'-terminated ssDNA. We biochemically reconstituted DNA end resection using purified human proteins: Bloom helicase (BLM); DNA2 helicase/nuclease; Exonuclease 1 (EXO1); the complex comprising MRE11, RAD50, and NBS1 (MRN); and Replication protein A (RPA). Resection occurs via two routes. In one, BLM and DNA2 physically and specifically interact to resect DNA in a process that is ATP-dependent and requires BLM helicase and DNA2 nuclease functions. RPA is essential for both DNA unwinding by BLM and enforcing 5' → 3' resection polarity by DNA2. MRN accelerates processing by recruiting BLM to the end. In the other, EXO1 resects the DNA and is stimulated by BLM, MRN, and RPA. BLM increases the affinity of EXO1 for ends, and MRN recruits and enhances the processivity of EXO1. Our results establish two of the core machineries that initiate recombinational DNA repair in human cells.

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Published In

Genes Dev

DOI

EISSN

1549-5477

Publication Date

February 15, 2011

Volume

25

Issue

4

Start / End Page

350 / 362

Location

United States

Related Subject Headings

  • Replication Protein A
  • RecQ Helicases
  • Protein Binding
  • Nuclear Proteins
  • Multiprotein Complexes
  • Models, Biological
  • MRE11 Homologue Protein
  • In Vitro Techniques
  • Humans
  • Exodeoxyribonucleases
 

Citation

APA
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Nimonkar, A. V., Genschel, J., Kinoshita, E., Polaczek, P., Campbell, J. L., Wyman, C., … Kowalczykowski, S. C. (2011). BLM-DNA2-RPA-MRN and EXO1-BLM-RPA-MRN constitute two DNA end resection machineries for human DNA break repair. Genes Dev, 25(4), 350–362. https://doi.org/10.1101/gad.2003811
Nimonkar, Amitabh V., Jochen Genschel, Eri Kinoshita, Piotr Polaczek, Judith L. Campbell, Claire Wyman, Paul Modrich, and Stephen C. Kowalczykowski. “BLM-DNA2-RPA-MRN and EXO1-BLM-RPA-MRN constitute two DNA end resection machineries for human DNA break repair.Genes Dev 25, no. 4 (February 15, 2011): 350–62. https://doi.org/10.1101/gad.2003811.
Nimonkar AV, Genschel J, Kinoshita E, Polaczek P, Campbell JL, Wyman C, et al. BLM-DNA2-RPA-MRN and EXO1-BLM-RPA-MRN constitute two DNA end resection machineries for human DNA break repair. Genes Dev. 2011 Feb 15;25(4):350–62.
Nimonkar, Amitabh V., et al. “BLM-DNA2-RPA-MRN and EXO1-BLM-RPA-MRN constitute two DNA end resection machineries for human DNA break repair.Genes Dev, vol. 25, no. 4, Feb. 2011, pp. 350–62. Pubmed, doi:10.1101/gad.2003811.
Nimonkar AV, Genschel J, Kinoshita E, Polaczek P, Campbell JL, Wyman C, Modrich P, Kowalczykowski SC. BLM-DNA2-RPA-MRN and EXO1-BLM-RPA-MRN constitute two DNA end resection machineries for human DNA break repair. Genes Dev. 2011 Feb 15;25(4):350–362.

Published In

Genes Dev

DOI

EISSN

1549-5477

Publication Date

February 15, 2011

Volume

25

Issue

4

Start / End Page

350 / 362

Location

United States

Related Subject Headings

  • Replication Protein A
  • RecQ Helicases
  • Protein Binding
  • Nuclear Proteins
  • Multiprotein Complexes
  • Models, Biological
  • MRE11 Homologue Protein
  • In Vitro Techniques
  • Humans
  • Exodeoxyribonucleases