Recombinational repair of gaps in DNA is asymmetric in Ustilago maydis and can be explained by a migrating D-loop model.

TitleRecombinational repair of gaps in DNA is asymmetric in Ustilago maydis and can be explained by a migrating D-loop model.
Publication TypeJournal Article
Year of Publication1996
AuthorsFerguson DO, Holloman WK
JournalProc Natl Acad Sci U S A
Date Published1996 May 28
KeywordsBase Sequence, Crossing Over, Genetic, DNA Primers, DNA Repair, DNA, Fungal, Genes, Fungal, Models, Genetic, Molecular Sequence Data, Polymerase Chain Reaction, Polymorphism, Restriction Fragment Length, Promoter Regions, Genetic, Recombination, Genetic, Restriction Mapping, Ustilago

Recombinational repair of double-stranded DNA gaps was investigated in Ustilago maydis. The experimental system was designed for analysis of repair of an autonomously replicating plasmid containing a cloned gene disabled by an internal deletion. It was discovered that crossing over rarely accompanied gap repair. The strong bias against crossing over was observed in three different genes regardless of gap size. These results indicate that gap repair in U. maydis is unlikely to proceed by the mechanism envisioned in the double-stranded break repair model of recombination, which was developed to account for recombination in Saccharomyces cerevisiae. Experiments aimed at exploring processing of DNA ends were performed to gain understanding of the mechanism responsible for the observed bias. A heterologous insert placed within a gap in the coding sequence of two different marker genes strongly inhibited repair if the DNA was cleaved at the promoter-proximal junction joining the insert and coding sequence but had little effect on repair if the DNA was cleaved at the promoter-distal junction. Gene conversion of plasmid restriction fragment length polymorphism markers engineered in sequences flanking both sides of a gap accompanied repair but was directionally biased. These results are interpreted to mean that the DNA ends flanking a gap are subject to different types of processing. A model featuring a single migrating D-loop is proposed to explain the bias in gap repair outcome based on the observed asymmetry in processing the DNA ends.

Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID8643590
PubMed Central IDPMC39261
Grant ListGM42482 / GM / NIGMS NIH HHS / United States

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