title: Mechanisms of nonhomologous recombination in mammalian cells.
abstract: The primary mechanism of nonhomologous recombination in transfected DNA involves breakage followed by end joining. To probe the joining step in more detail, linear simian virus 40 genomes with mismatched ends were transfected into cultured monkey cells, and individual viable recombinants were analyzed. The transfected genomes carried mismatched ends as a result of cleavage with two restriction enzymes, the recognition sites of which are located in the intron of the gene encoding the T antigen. Because the T antigen gene was split by this cleavage, the transfected genomes were inert until activated by cell-mediated end joining. Clonal descendants of the original recombinants were isolated from 122 plaques and were grouped into four classes based on the electrophoretic mobility of the junction fragment. The structures of representative junctions were determined by nucleotide sequencing. The spectrum of nonhomologous junctions analyzed here along with a large number of previously reported junctions suggest that there are two mechanisms for the linkage of DNA molecules: (i) direct ligation of ends and (ii) repair synthesis primed by terminal homologies of a few nucleotides. A paired-priming model of nonhomologous recombination is discussed.
mesh_terms: D000818:Animals; D000957:Antigens, Viral, Tumor / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D004260:DNA Repair; D004279:DNA, Viral / Q000235:genetics; D011995:Recombination, Genetic; D013539:Simian virus 40 / Q000235:genetics
chemical_list: D000957:Antigens, Viral, Tumor; D004279:DNA, Viral