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DNA binding, annealing, and strand exchange activities of Brh2 protein from Ustilago maydis.

TitleDNA binding, annealing, and strand exchange activities of Brh2 protein from Ustilago maydis.
Publication TypeJournal Article
Year of Publication2007
AuthorsMazloum N, Zhou Q, Holloman WK
JournalBiochemistry
Volume46
Issue24
Pagination7163-73
Date Published2007 Jun 19
ISSN0006-2960
KeywordsApoptosis Regulatory Proteins, Base Sequence, BRCA2 Protein, DNA, DNA Repair, DNA-Binding Proteins, Fungal Proteins, Humans, Kinetics, Rad51 Recombinase, Recombinant Proteins, Replication Protein A, Ustilago
Abstract

Brh2 is the Ustilago maydis ortholog of the BRCA2 tumor suppressor. It functions in repair of DNA by homologous recombination by controlling the action of Rad51. A critical aspect in the control appears to be the recruitment of Rad51 to single-stranded DNA regions exposed as lesions after damage or following a disturbance in DNA synthesis. In previous experimentation, Brh2 was shown to nucleate formation of the Rad51 nucleoprotein filament that becomes the active element in promoting homologous pairing and DNA strand exchange. Nucleation was found to be initiated at junctions of double-stranded and single-stranded DNA. Here we investigated the DNA binding specificity of Brh2 in more detail using oligonucleotide substrates. We observed that Brh2 prefers partially duplex structures with single-stranded branches, flaps, or D-loops. We found also that Brh2 has an inherent ability to promote DNA annealing and strand exchange reactions on free as well as RPA-coated substrates. Unlike Rad51, Brh2 was able to promote DNA strand exchange when preincubated with double-stranded DNA. These findings raise the notion that Brh2 may have roles in homologous recombination beyond the previously established Rad51 mediator activity.

DOI10.1021/bi700399m
Alternate JournalBiochemistry
PubMed ID17523678
PubMed Central IDPMC2553723
Grant ListR01 GM042482 / GM / NIGMS NIH HHS / United States
R01 GM042482-16 / GM / NIGMS NIH HHS / United States
R01 GM079859 / GM / NIGMS NIH HHS / United States
R01 GM079859-01A1 / GM / NIGMS NIH HHS / United States

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