CHARACTERIZATION OF NONCONSERVATIVE HOMOLOGOUS JUNCTIONS IN MAMMALIAN-CELLS

被引:15
作者
DESAUTELS, L
BROUILLETTE, S
WALLENBURG, J
BELMAAZA, A
GUSEW, N
TRUDEL, P
CHARTRAND, P
机构
[1] CANADIAN RED CROSS SOC,BLOOD SERV,MONTREAL CTR,3131 SHERBROOKE ST E,MONTREAL H1W 1B2,QUEBEC,CANADA
[2] UNIV MONTREAL,DEPT MICROBIOL & IMMUNOL,MONTREAL H3C 3J7,QUEBEC,CANADA
关键词
D O I
10.1128/MCB.10.12.6613
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Homologous recombination in mammalian cells between extrachromosomal molecules, as well as between episomes and chromosomes, can be mediated by a nonconservative mechanism. It has been proposed that the key steps in this process are the generation (by double-strand cleavage) of overlapping homologous ends, the creation of complementary single-strand ends (either by strand-specific exonuclease degradation or by unwinding of the DNA helix), and finally the creation of heteroduplex DNA by the annealing of the single-strand ends. We have analyzed in detail the structure of nonconservative homologous junctions and determined the contribution of each end to the formation of the junction. We have also analyzed multiple descendants from single recombination events. Two types of junctions were found. The majority (90%) of the junctions were characterized by a single crossover site. These crossover sites were distributed randomly throughout the junction. The remaining 10% of the junctions had mosaic patterns of parental markers. Furthermore, in 9 of 10 cases, multiple descendants from a single recombination event were identical. Thus, it appears that in most cases few parental markers were involved in junction formation. This finding suggests that nonconservative homologous junctions are mediated mainly by short heteroduplexes of a few hundred base pairs or less. These results are discussed in terms of the current models of nonconservative homologous recombination.
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页码:6613 / 6618
页数:6
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