Colocalization of multiple DNA double-strand breaks at a single Rad52 repair centre

TY - JOUR

T1 - Colocalization of multiple DNA double-strand breaks at a single Rad52 repair centre

AU - Lisby, Michael

AU - Mortensen, Uffe H

AU - Rothstein, Rodney

PY - 2003/6

Y1 - 2003/6

N2 - DNA double-strand break repair (DSBR) is an essential process for preserving genomic integrity in all organisms. To investigate this process at the cellular level, we engineered a system of fluorescently marked DNA double-strand breaks (DSBs) in the yeast Saccharomyces cerevisiae to visualize in vivo DSBR in single cells. Using this system, we demonstrate for the first time that Rad52 DNA repair foci and DSBs colocalize. Time-lapse microscopy reveals that the relocalization of Rad52 protein into a focal assembly is a rapid and reversible process. In addition, analysis of DNA damage checkpoint-deficient cells provides direct evidence for coordination between DNA repair and subsequent release from checkpoint arrest. Finally, analyses of cells experiencing multiple DSBs demonstrate that Rad52 foci are centres of DNA repair capable of simultaneously recruiting more than one DSB.

AB - DNA double-strand break repair (DSBR) is an essential process for preserving genomic integrity in all organisms. To investigate this process at the cellular level, we engineered a system of fluorescently marked DNA double-strand breaks (DSBs) in the yeast Saccharomyces cerevisiae to visualize in vivo DSBR in single cells. Using this system, we demonstrate for the first time that Rad52 DNA repair foci and DSBs colocalize. Time-lapse microscopy reveals that the relocalization of Rad52 protein into a focal assembly is a rapid and reversible process. In addition, analysis of DNA damage checkpoint-deficient cells provides direct evidence for coordination between DNA repair and subsequent release from checkpoint arrest. Finally, analyses of cells experiencing multiple DSBs demonstrate that Rad52 foci are centres of DNA repair capable of simultaneously recruiting more than one DSB.

KW - Bacterial Proteins

KW - DNA Damage

KW - DNA Repair

KW - DNA, Fungal

KW - DNA-Binding Proteins

KW - Deoxyribonucleases, Type II Site-Specific

KW - Escherichia coli Proteins

KW - G1 Phase

KW - G2 Phase

KW - Gamma Rays

KW - Genes, Insect

KW - Haploidy

KW - Lac Repressors

KW - Mitosis

KW - Rad52 DNA Repair and Recombination Protein

KW - Recombinant Fusion Proteins

KW - Repressor Proteins

KW - Saccharomyces cerevisiae

KW - Saccharomyces cerevisiae Proteins

KW - Time Factors

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

KW - Research Support, U.S. Gov't, P.H.S.

U2 - 10.1038/ncb997

DO - 10.1038/ncb997

M3 - Journal article

C2 - 12766777

SN - 1465-7392

VL - 5

SP - 572

EP - 577

JO - Nature Cell Biology

JF - Nature Cell Biology

IS - 6

ER -