RAD51-dependent break-induced replication in yeast.
A chromosome fragmentation assay was used to measure the efficiency and genetic control of break-induced replication (BIR) in Saccharomyces cerevisiae. Formation of a chromosome fragment by de novo telomere generation at one end of the linear vector and recombination-dependent replication of 100 kb of chromosomal sequences at the other end of the vector occurred at high frequency in wild-type strains. RAD51 was required for more than 95% of BIR events involving a single-end invasion and was essential when two BIR events were required for generation of a chromosome fragment. The similar genetic requirements for BIR and gene conversion suggest a common strand invasion intermediate in these two recombinational repair processes. Mutation of RAD50 or RAD59 conferred no significant defect in BIR in either RAD51 or rad51 strains. RAD52 was shown to be essential for BIR at unique chromosomal sequences, although rare recombination events were detected between the subtelomeric Y' repeats.
Department of Microbiology and Institute of Cancer Research, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
Allison P
AP
Davis
Lorraine S
LS
Symington
D000251:Adenosine Triphosphatases; D001483:Base Sequence; D019457:Chromosome Breakage; D015825:Chromosomes, Fungal / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D004265:DNA Helicases; D004260:DNA Repair; D045643:DNA Repair Enzymes; D004261:DNA Replication / Q000235:genetics / Q000502:physiology*; D004271:DNA, Fungal / Q000096:biosynthesis / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005800:Genes, Fungal; D005822:Genetic Vectors; D009154:Mutation; D051135:Rad51 Recombinase; D051721:Rad52 DNA Repair and Recombination Protein; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; C497197:RAD52 protein, S cerevisiae; C087191:RAD55 protein, S cerevisiae; D051721:Rad52 DNA Repair and Recombination Protein; D029701:Saccharomyces cerevisiae Proteins; C495151:RAD51 protein, S cerevisiae; D051135:Rad51 Recombinase; D000251:Adenosine Triphosphatases; C075468:RAD54 protein, S cerevisiae; C071451:RAD57 protein, S cerevisiae; D004265:DNA Helicases; D045643:DNA Repair Enzymes