Assembly of RecA-like recombinases: distinct roles for mediator proteins in mitosis and meiosis.
Members of the RecA family of recombinases from bacteriophage T4, Escherichia coli, yeast, and higher eukaryotes function in recombination as higher-order oligomers assembled on tracts of single-strand DNA (ssDNA). Biochemical studies have shown that assembly of recombinase involves accessory factors. These studies have identified a class of proteins, called recombination mediator proteins, that act by promoting assembly of recombinase on ssDNA tracts that are bound by ssDNA-binding protein (ssb). In the absence of mediators, ssb inhibits recombination reactions by competing with recombinase for DNA-binding sites. Here we briefly review mediated recombinase assembly and present results of new in vivo experiments. Immuno-double-staining experiments in Saccharomyces cerevisiae suggest that Rad51, the eukaryotic recombinase, can assemble at or near sites containing ssb (replication protein A, RPA) during the response to DNA damage, consistent with a need for mediator activity. Correspondingly, mediator gene mutants display defects in Rad51 assembly after DNA damage and during meiosis, although the requirements for assembly are distinct in the two cases. In meiosis, both Rad52 and Rad55/57 are required, whereas either Rad52 or Rad55/57 is sufficient to promote assembly of Rad51 in irradiated mitotic cells. Rad52 promotes normal amounts of Rad51 assembly in the absence of Rad55 at 30 degrees C but not 20 degrees C, accounting for the cold sensitivity of rad55 null mutants. Finally, we show that assembly of Rad51 is induced by radiation during S phase but not during G(1), consistent with the role of Rad51 in repairing the spontaneous damage that occurs during DNA replication.
Gasior
S L
SL
Department of Radiation and Cellular Oncology, University of Chicago, Chicago, IL 60637, USA.
Olivares
H
H
Ear
U
U
Hari
D M
DM
Weichselbaum
R
R
Bishop
D K
DK
D000251:Adenosine Triphosphatases; D000818:Animals; D004249:DNA Damage; D045643:DNA Repair Enzymes; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005057:Eukaryotic Cells; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism; D006801:Humans; D008540:Meiosis / Q000502:physiology*; D008938:Mitosis / Q000502:physiology*; D051135:Rad51 Recombinase; D011938:Rec A Recombinases / Q000378:metabolism; D011995:Recombination, Genetic; D051716:Replication Protein A; D029701:Saccharomyces cerevisiae Proteins
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C087191:RAD55 protein, S cerevisiae; C497167:RPA1 protein, human; D051716:Replication Protein A; D029701:Saccharomyces cerevisiae Proteins; C495153:RAD51 protein, human; D051135:Rad51 Recombinase; D011938:Rec A Recombinases; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V; D000251:Adenosine Triphosphatases; C071451:RAD57 protein, S cerevisiae; D045643:DNA Repair Enzymes