Microbiology and Immunology

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Approaches to Understanding the Mediator Function of Brh2 in Ustilago maydis.

TitleApproaches to Understanding the Mediator Function of Brh2 in Ustilago maydis.
Publication TypeJournal Article
Year of Publication2018
AuthorsZhou Q, Holloman WK, Kojic M
JournalMethods Enzymol
Volume600
Pagination513-525
Date Published2018
ISSN1557-7988
Abstract

Primary components of the homologous recombination pathway in eukaryotes include Rad51 whose function is to search for DNA sequence homology and promote strand exchange, its mediator BRCA2, and Dss1, a key regulator of BRCA2. We seek to understand the role of BRCA2 in governing the activity of Rad51 and to learn how BRCA2 function is regulated by Dss1. We use the microbe Ustilago maydis as a model system for experimentation because it has a well-conserved BRCA2-homolog, Brh2, and is amenable to biochemical and molecular genetic manipulations and analysis. The powerful attributes of this system open the way for gaining insight into BRCA2's molecular mechanism through avenues not immediately approachable in the vertebrate systems. Here we provide protocols for preparing Brh2, Dss1, and Rad51 as reagents for use in biochemical assays to monitor function and present methods for transposon-based mutational analysis of Brh2 for use in genetic dissection of function.

DOI10.1016/bs.mie.2017.11.019
Alternate JournalMeth. Enzymol.
PubMed ID29458772

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