The role of BRCA2 in replication-coupled DNA interstrand cross-link repair in vitro

Lubos Cipak, Norifumi Watanabe, Tadayoshi Bessho

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Using a defined substrate DNA with a single psoralen interstrand cross-link (ICL), we studied the molecular mechanism of human ICL repair. In vitro ICL repair by human extracts is dependent on replication and is a largely error-free process. Extracts from a human BRCA2-defective mutant cell line, CAPAN-1, are severely compromised in ICL repair. Specifically, 'unhooked' but not fully repaired products accumulate in the reaction with CAPAN-1, and transient expression of BRCA2 in CAPAN-1 restores repair activity. Together, these results reveal that BRCA2 participates in repair of replication-mediated double-strand breaks generated when replication forks encounter ICLs. We also show that nucleotide excision repair is essential for the removal of the lesion left behind on one strand after unhooking. This study provides new mechanistic insights into the repair of ICLs in human cells.

Original languageEnglish (US)
Pages (from-to)729-733
Number of pages5
JournalNature Structural and Molecular Biology
Volume13
Issue number8
DOIs
StatePublished - Aug 1 2006

Fingerprint

DNA Replication
Ficusin
DNA Repair
Cell Line
In Vitro Techniques
DNA

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

The role of BRCA2 in replication-coupled DNA interstrand cross-link repair in vitro. / Cipak, Lubos; Watanabe, Norifumi; Bessho, Tadayoshi.

In: Nature Structural and Molecular Biology, Vol. 13, No. 8, 01.08.2006, p. 729-733.

Research output: Contribution to journalArticle

@article{6fc260da2d5f4983991604c02c8def35,
title = "The role of BRCA2 in replication-coupled DNA interstrand cross-link repair in vitro",
abstract = "Using a defined substrate DNA with a single psoralen interstrand cross-link (ICL), we studied the molecular mechanism of human ICL repair. In vitro ICL repair by human extracts is dependent on replication and is a largely error-free process. Extracts from a human BRCA2-defective mutant cell line, CAPAN-1, are severely compromised in ICL repair. Specifically, 'unhooked' but not fully repaired products accumulate in the reaction with CAPAN-1, and transient expression of BRCA2 in CAPAN-1 restores repair activity. Together, these results reveal that BRCA2 participates in repair of replication-mediated double-strand breaks generated when replication forks encounter ICLs. We also show that nucleotide excision repair is essential for the removal of the lesion left behind on one strand after unhooking. This study provides new mechanistic insights into the repair of ICLs in human cells.",
author = "Lubos Cipak and Norifumi Watanabe and Tadayoshi Bessho",
year = "2006",
month = "8",
day = "1",
doi = "10.1038/nsmb1120",
language = "English (US)",
volume = "13",
pages = "729--733",
journal = "Nature Structural and Molecular Biology",
issn = "1545-9993",
publisher = "Nature Publishing Group",
number = "8",

}

TY - JOUR

T1 - The role of BRCA2 in replication-coupled DNA interstrand cross-link repair in vitro

AU - Cipak, Lubos

AU - Watanabe, Norifumi

AU - Bessho, Tadayoshi

PY - 2006/8/1

Y1 - 2006/8/1

N2 - Using a defined substrate DNA with a single psoralen interstrand cross-link (ICL), we studied the molecular mechanism of human ICL repair. In vitro ICL repair by human extracts is dependent on replication and is a largely error-free process. Extracts from a human BRCA2-defective mutant cell line, CAPAN-1, are severely compromised in ICL repair. Specifically, 'unhooked' but not fully repaired products accumulate in the reaction with CAPAN-1, and transient expression of BRCA2 in CAPAN-1 restores repair activity. Together, these results reveal that BRCA2 participates in repair of replication-mediated double-strand breaks generated when replication forks encounter ICLs. We also show that nucleotide excision repair is essential for the removal of the lesion left behind on one strand after unhooking. This study provides new mechanistic insights into the repair of ICLs in human cells.

AB - Using a defined substrate DNA with a single psoralen interstrand cross-link (ICL), we studied the molecular mechanism of human ICL repair. In vitro ICL repair by human extracts is dependent on replication and is a largely error-free process. Extracts from a human BRCA2-defective mutant cell line, CAPAN-1, are severely compromised in ICL repair. Specifically, 'unhooked' but not fully repaired products accumulate in the reaction with CAPAN-1, and transient expression of BRCA2 in CAPAN-1 restores repair activity. Together, these results reveal that BRCA2 participates in repair of replication-mediated double-strand breaks generated when replication forks encounter ICLs. We also show that nucleotide excision repair is essential for the removal of the lesion left behind on one strand after unhooking. This study provides new mechanistic insights into the repair of ICLs in human cells.

UR - http://www.scopus.com/inward/record.url?scp=33746811317&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33746811317&partnerID=8YFLogxK

U2 - 10.1038/nsmb1120

DO - 10.1038/nsmb1120

M3 - Article

C2 - 16845393

AN - SCOPUS:33746811317

VL - 13

SP - 729

EP - 733

JO - Nature Structural and Molecular Biology

JF - Nature Structural and Molecular Biology

SN - 1545-9993

IS - 8

ER -