Nuclear dynamics of RAD52 group homologous recombination proteins in response to DNA damage

Recombination between homologous DNA molecules is essential for the proper maintenance and duplication of the genome, and for the repair of exogenously induced DNA damage such as double‐strand breaks. Homologous recombination requires the RAD52 group proteins, including Rad51, Rad52 and Rad54. Upon treatment of mammalian cells with ionizing radiation, these proteins accumulate into foci at sites of DNA damage induction. We show that these foci are dynamic structures of which Rad51 is a stably associated core component, whereas Rad52 and Rad54 rapidly and reversibly interact with the structure. Furthermore, we show that the majority of the proteins are not part of the same multi‐protein complex in the absence of DNA damage. Executing DNA transactions through dynamic multi‐protein complexes, rather than stable holo‐complexes, allows flexibility. In the case of DNA repair, for example, it will facilitate cross‐talk between different DNA repair pathways and coupling to other DNA transactions, such as replication.

[1]  R. Kanaar,et al.  Brca2 (XRCC11) Deficiency Results in Radioresistant DNA Synthesis and a Higher Frequency of Spontaneous Deletions , 2002, Molecular and Cellular Biology.

[2]  Howard J. Worman,et al.  Nuclear Membrane Dynamics and Reassembly in Living Cells: Targeting of an Inner Nuclear Membrane Protein in Interphase and Mitosis , 1997, The Journal of cell biology.

[3]  P. Berg,et al.  Complex formation in yeast double-strand break repair: participation of Rad51, Rad52, Rad55, and Rad57 proteins. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[4]  A. Houtsmuller,et al.  Action of DNA repair endonuclease ERCC1/XPF in living cells. , 1999, Science.

[5]  K. Knight,et al.  The hRad51 and RecA proteins show significant differences in cooperative binding to single-stranded DNA. , 1999, Journal of molecular biology.

[6]  P. Baumann,et al.  Role of the human RAD51 protein in homologous recombination and double-stranded-break repair. , 1998, Trends in biochemical sciences.

[7]  P. Sung,et al.  Recombination factors of Saccharomyces cerevisiae. , 2000, Mutation research.

[8]  D. Ward,et al.  Nuclear foci of mammalian Rad51 recombination protein in somatic cells after DNA damage and its localization in synaptonemal complexes. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[9]  T. Pederson,et al.  Protein Mobility within the Nucleus—What Are the Right Moves? , 2001, Cell.

[10]  R. Kolodner,et al.  Links between replication, recombination and genome instability in eukaryotes. , 2000, Trends in biochemical sciences.

[11]  R. Rothstein,et al.  Rad52 forms DNA repair and recombination centers during S phase , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[12]  T. Cremer,et al.  Chromosome territories, nuclear architecture and gene regulation in mammalian cells , 2001, Nature Reviews Genetics.

[13]  S C West,et al.  Role of BRCA2 in control of the RAD51 recombination and DNA repair protein. , 2001, Molecular cell.

[14]  D. Schild,et al.  Mutants of the Five Rad51 Paralogs Recombinational Repair in Knockout Chromosome Instability and Defective , 2022 .

[15]  T Misteli,et al.  Protein dynamics: implications for nuclear architecture and gene expression. , 2001, Science.

[16]  S. West,et al.  Gross chromosomal rearrangements and genetic exchange between nonhomologous chromosomes following BRCA2 inactivation. , 2000, Genes & development.

[17]  R. Weichselbaum,et al.  Xrcc3 Is Required for Assembly of Rad51 Complexes in Vivo * , 1998, The Journal of Biological Chemistry.

[18]  J. Hoeijmakers,et al.  Mouse Rad54 affects DNA conformation and DNA-damage-induced Rad51 foci formation , 1999, Current Biology.

[19]  C. Wilson,et al.  XRCC2 Is a Nuclear RAD51-like Protein Required for Damage-dependent RAD51 Focus Formation without the Need for ATP Binding* , 2001, The Journal of Biological Chemistry.

[20]  T. Haaf,et al.  Nuclear foci of mammalian recombination proteins are located at single-stranded DNA regions formed after DNA damage. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[21]  R. Kanaar,et al.  Homologous recombination: from model organisms to human disease , 2001, Genome Biology.

[22]  S. West,et al.  Binding of double-strand breaks in DNA by human Rad52 protein , 1999, Nature.

[23]  J. Albala,et al.  The Rad51 Paralog Rad51B Promotes Homologous Recombinational Repair , 2000, Molecular and Cellular Biology.

[24]  J. Lamerdin,et al.  XRCC2 and XRCC3, new human Rad51-family members, promote chromosome stability and protect against DNA cross-links and other damages. , 1998, Molecular cell.

[25]  D. Ward,et al.  Interaction of human recombination proteins Rad51 and Rad54. , 1997, Nucleic acids research.

[26]  N. Kamada,et al.  Rad51 Accumulation at Sites of DNA Damage and in Postreplicative Chromatin , 2000, The Journal of cell biology.

[27]  Adriaan B. Houtsmuller,et al.  Macromolecular dynamics in living cell nuclei revealed by fluorescence redistribution after photobleaching , 2001, Histochemistry and Cell Biology.

[28]  N. Maizels,et al.  Coordinated response of mammalian Rad51 and Rad52 to DNA damage , 2000, EMBO reports.

[29]  L. Symington,et al.  Copyright � 1995, American Society for Microbiology Functional Differences and Interactions among the Putative , 1995 .

[30]  J. Hoeijmakers,et al.  Disruption of Mouse RAD54 Reduces Ionizing Radiation Resistance and Homologous Recombination , 1997, Cell.

[31]  R. Kanaar,et al.  Repair of DNA interstrand cross-links. , 2001, Mutation research.

[32]  S. West,et al.  The human Rad52 protein exists as a heptameric ring , 2000, Current Biology.

[33]  D. Baltimore,et al.  Radiation-induced Assembly of Rad51 and Rad52 Recombination Complex Requires ATM and c-Abl* , 1999, The Journal of Biological Chemistry.

[34]  E. Stelzer,et al.  Photobleaching GFP reveals protein dynamics inside live cells. , 1999, Trends in cell biology.

[35]  Richard Cartwright,et al.  The XRCC2 DNA repair gene from human and mouse encodes a novel member of the recA/RAD51 family , 1998, Nucleic Acids Res..

[36]  J. Haber,et al.  Multiple Pathways of Recombination Induced by Double-Strand Breaks in Saccharomyces cerevisiae , 1999, Microbiology and Molecular Biology Reviews.

[37]  T. Misteli,et al.  High mobility of proteins in the mammalian cell nucleus , 2000, Nature.