Approaches to Understanding the Mediator Function of Brh2 in Ustilago maydis
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.
Source:
Mechanisms of Dna Recombination and Genome Rearrangements: Methods To Study Homologous Recombination, 2018, 600, 513-525Publisher:
- Elsevier Academic Press Inc, San Diego
Funding / projects:
- The Role of Transcription Factors and Small RNAs in Abiotic Stress Response in Plants and Genetic Diversity of Plant Species Important for Agriculture and Biotechnology (RS-MESTD-Basic Research (BR or ON)-173005)
DOI: 10.1016/bs.mie.2017.11.019
ISSN: 0076-6879
PubMed: 29458772
WoS: 000452357600021
Scopus: 2-s2.0-85041110383
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Institution/Community
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Zhou, Qingwen AU - Holloman, William K. AU - Kojić, Milorad PY - 2018 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/1138 AB - 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. PB - Elsevier Academic Press Inc, San Diego T2 - Mechanisms of Dna Recombination and Genome Rearrangements: Methods To Study Homologous Recombination T1 - Approaches to Understanding the Mediator Function of Brh2 in Ustilago maydis EP - 525 SP - 513 VL - 600 DO - 10.1016/bs.mie.2017.11.019 ER -
@article{ author = "Zhou, Qingwen and Holloman, William K. and Kojić, Milorad", year = "2018", 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.", publisher = "Elsevier Academic Press Inc, San Diego", journal = "Mechanisms of Dna Recombination and Genome Rearrangements: Methods To Study Homologous Recombination", title = "Approaches to Understanding the Mediator Function of Brh2 in Ustilago maydis", pages = "525-513", volume = "600", doi = "10.1016/bs.mie.2017.11.019" }
Zhou, Q., Holloman, W. K.,& Kojić, M.. (2018). Approaches to Understanding the Mediator Function of Brh2 in Ustilago maydis. in Mechanisms of Dna Recombination and Genome Rearrangements: Methods To Study Homologous Recombination Elsevier Academic Press Inc, San Diego., 600, 513-525. https://doi.org/10.1016/bs.mie.2017.11.019
Zhou Q, Holloman WK, Kojić M. Approaches to Understanding the Mediator Function of Brh2 in Ustilago maydis. in Mechanisms of Dna Recombination and Genome Rearrangements: Methods To Study Homologous Recombination. 2018;600:513-525. doi:10.1016/bs.mie.2017.11.019 .
Zhou, Qingwen, Holloman, William K., Kojić, Milorad, "Approaches to Understanding the Mediator Function of Brh2 in Ustilago maydis" in Mechanisms of Dna Recombination and Genome Rearrangements: Methods To Study Homologous Recombination, 600 (2018):513-525, https://doi.org/10.1016/bs.mie.2017.11.019 . .