Slipped misalignment mechanisms of deletion formation: in vivo susceptibility to nucleases.
Misalignment of repeated sequences during DNA replication can lead to deletions or duplications in genomic DNA. In Escherichia coli, such genetic rearrangements can occur at high frequencies, independent of the RecA-homologous recombination protein, and are sometimes associated with sister chromosome exchange (SCE). Two mechanisms for RecA-independent genetic rearrangements have been proposed: simple replication misalignment of the nascent strand and its template and SCE-associated misalignment involving both nascent strands. We examined the influence of the 3' exonuclease of DNA polymerase III and exonuclease I on deletion via these mechanisms in vivo. Because mutations in these exonucleases stimulate tandem repeat deletion, we conclude that displaced 3' ends are a common intermediate in both mechanisms of slipped misalignments. Our results also confirm the notion that two distinct mechanisms contribute to slipped misalignments: simple replication misalignment events are sensitive to DNA polymerase III exonuclease, whereas SCE-associated events are sensitive to exonuclease I. If heterologies are present between repeated sequences, the mismatch repair system dependent on MutS and MutH aborts potential deletion events via both mechanisms. Our results suggest that simple slipped misalignment and SCE-associated misalignment intermediates are similarly susceptible to destruction by the mismatch repair system.
Bzymek
M
M
Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, Massachusetts 02454-9110, USA.
Saveson
C J
CJ
Feschenko
V V
VV
Lovett
S T
ST
D004258:DNA Polymerase III / Q000378:metabolism*; D004261:DNA Replication; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D005092:Exonucleases / Q000378:metabolism*; D015321:Gene Rearrangement; D008957:Models, Genetic; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D012091:Repetitive Sequences, Nucleic Acid; D016415:Sequence Alignment; D017384:Sequence Deletion; D013698:Templates, Genetic
D004269:DNA, Bacterial; D004258:DNA Polymerase III; D005090:Exodeoxyribonucleases; D005092:Exonucleases; C028450:exodeoxyribonuclease I; C022650:spleen exonuclease