DNA end-joining driven by microhomologies catalyzed by nuclear extracts

被引:3
作者
Boán, F [1 ]
Blanco, MG [1 ]
Barros, P [1 ]
Gómez-Márquez, J [1 ]
机构
[1] Univ Santiago de Compostela, Fac Biol, Dept Bioquim & Biol Mol, E-15782 Santiago De Compostela, Spain
关键词
double-strand break; microhomologies; minisatellite MsH42; non-homologous end-joining; recombination;
D O I
10.1515/BC.2006.035
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In a previous work we used an in vitro system for the generation and analysis of double-strand breaks (DSBs) using nuclear extracts from rat testes as a source of DSB activity. Since the recombination process can be triggered by the formation of DSB, in the present study we developed a strategy to isolate and characterize recombinant molecules using the same in vitro system. Our results indicate that the mechanism for the formation of recombinants was non-homologous end-joining driven by microhomologies. The procedure described here represents an alternative to investigate the mechanisms of DNA end-joining and other forms of DNA repair.
引用
收藏
页码:263 / 267
页数:5
相关论文
共 29 条
[1]  
Ausubel F.M., 1994, CURRENT PROTOCOLS MO
[2]   DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis [J].
Bartkova, J ;
Horejsi, Z ;
Koed, K ;
Krämer, A ;
Tort, F ;
Zieger, K ;
Guldberg, P ;
Sehested, M ;
Nesland, JM ;
Lukas, C ;
Orntoft, T ;
Lukas, J ;
Bartek, J .
NATURE, 2005, 434 (7035) :864-870
[3]   DNA end-joining catalyzed by human cell-free extracts [J].
Baumann, P ;
West, SC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (24) :14066-14070
[4]   Generation of DNA double-strand breaks by two independent enzymatic activities in nuclear extracts [J].
Blanco, MG ;
Boán, F ;
Barros, P ;
Castaño, JG ;
Gómez-Márquez, J .
JOURNAL OF MOLECULAR BIOLOGY, 2005, 351 (05) :995-1006
[5]   A paradox in the in vitro end-joining assays [J].
Blanco, MG ;
Boán, F ;
Gómez-Márquez, J .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (25) :26797-26801
[6]   Molecular characterization of a new human minisatellite that is able to form single-stranded loops in vitro and is recognized by nuclear proteins [J].
Boan, F ;
Gonzalez, AI ;
Rodriguez, JM ;
GomezMarquez, J .
FEBS LETTERS, 1997, 418 (03) :251-257
[7]   Recombination analysis of the human minisatellite MsH42 suggests the existence of two distinct pathways for initiation and resolution of recombination at MsH42 in rat testes nuclear extracts [J].
Boán, F ;
Rodríguez, JM ;
Mouriño, S ;
Blanco, MG ;
Viñas, A ;
Sánchez, L ;
Gómez-Márquez, J .
BIOCHEMISTRY, 2002, 41 (07) :2166-2176
[8]   A non-hypervariable human minisatellite strongly stimulates in vitro intramolecular homologous recombination [J].
Boán, F ;
Rodríguez, JM ;
Gómez-Márquez, J .
JOURNAL OF MOLECULAR BIOLOGY, 1998, 278 (03) :499-505
[9]   Minisatellite instability and germline mutation [J].
Bois, P ;
Jeffreys, AJ .
CELLULAR AND MOLECULAR LIFE SCIENCES, 1999, 55 (12) :1636-1648
[10]   Saccharomyces cerevisiae Ku70 potentiates illegitimate DNA double-strand break repair and serves as a barrier to error-prone DNA repair pathways [J].
Boulton, SJ ;
Jackson, SP .
EMBO JOURNAL, 1996, 15 (18) :5093-5103