Tags

Type your tag names separated by a space and hit enter

Functional analyses of the C-terminal half of the Saccharomyces cerevisiae Rad52 protein.
Nucleic Acids Res. 2014 Jan; 42(2):941-51.NA

Abstract

The Saccharomyces cerevisiae Rad52 protein is essential for efficient homologous recombination (HR). An important role of Rad52 in HR is the loading of Rad51 onto replication protein A-coated single-stranded DNA (ssDNA), which is referred to as the recombination mediator activity. In vitro, Rad52 displays additional activities, including self-association, DNA binding and ssDNA annealing. Although Rad52 has been a subject of extensive genetic, biochemical and structural studies, the mechanisms by which these activities are coordinated in the various roles of Rad52 in HR remain largely unknown. In the present study, we found that an isolated C-terminal half of Rad52 disrupted the Rad51 oligomer and formed a heterodimeric complex with Rad51. The Rad52 fragment inhibited the binding of Rad51 to double-stranded DNA, but not to ssDNA. The phenylalanine-349 and tyrosine-409 residues present in the C-terminal half of Rad52 were critical for the interaction with Rad51, the disruption of Rad51 oligomers, the mediator activity of the full-length protein and for DNA repair in vivo in the presence of methyl methanesulfonate. Our studies suggested that phenylalanine-349 and tyrosine-409 are key residues in the C-terminal half of Rad52 and probably play an important role in the mediator activity.

Authors+Show Affiliations

Department of Interdisciplinary Science and Engineering, Program in Chemistry and Life Science, School of Science and Engineering, Meisei University, 2-1-1 Hodokubo, Hino-shi, Tokyo 191-8506, Japan, Department of Applied Biological Science, Nihon University College of Bioresource Sciences, Fujisawa-shi, Kanagawa 252-0880, Japan, Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8480, Japan and Cellular and Molecular Biology Laboratory, RIKEN, Wako-shi, Saitama 351-0198, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24163251

Citation

Kagawa, Wataru, et al. "Functional Analyses of the C-terminal Half of the Saccharomyces Cerevisiae Rad52 Protein." Nucleic Acids Research, vol. 42, no. 2, 2014, pp. 941-51.
Kagawa W, Arai N, Ichikawa Y, et al. Functional analyses of the C-terminal half of the Saccharomyces cerevisiae Rad52 protein. Nucleic Acids Res. 2014;42(2):941-51.
Kagawa, W., Arai, N., Ichikawa, Y., Saito, K., Sugiyama, S., Saotome, M., Shibata, T., & Kurumizaka, H. (2014). Functional analyses of the C-terminal half of the Saccharomyces cerevisiae Rad52 protein. Nucleic Acids Research, 42(2), 941-51. https://doi.org/10.1093/nar/gkt986
Kagawa W, et al. Functional Analyses of the C-terminal Half of the Saccharomyces Cerevisiae Rad52 Protein. Nucleic Acids Res. 2014;42(2):941-51. PubMed PMID: 24163251.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Functional analyses of the C-terminal half of the Saccharomyces cerevisiae Rad52 protein. AU - Kagawa,Wataru, AU - Arai,Naoto, AU - Ichikawa,Yuichi, AU - Saito,Kengo, AU - Sugiyama,Shusei, AU - Saotome,Mika, AU - Shibata,Takehiko, AU - Kurumizaka,Hitoshi, Y1 - 2013/10/25/ PY - 2013/10/29/entrez PY - 2013/10/29/pubmed PY - 2014/3/26/medline SP - 941 EP - 51 JF - Nucleic acids research JO - Nucleic Acids Res VL - 42 IS - 2 N2 - The Saccharomyces cerevisiae Rad52 protein is essential for efficient homologous recombination (HR). An important role of Rad52 in HR is the loading of Rad51 onto replication protein A-coated single-stranded DNA (ssDNA), which is referred to as the recombination mediator activity. In vitro, Rad52 displays additional activities, including self-association, DNA binding and ssDNA annealing. Although Rad52 has been a subject of extensive genetic, biochemical and structural studies, the mechanisms by which these activities are coordinated in the various roles of Rad52 in HR remain largely unknown. In the present study, we found that an isolated C-terminal half of Rad52 disrupted the Rad51 oligomer and formed a heterodimeric complex with Rad51. The Rad52 fragment inhibited the binding of Rad51 to double-stranded DNA, but not to ssDNA. The phenylalanine-349 and tyrosine-409 residues present in the C-terminal half of Rad52 were critical for the interaction with Rad51, the disruption of Rad51 oligomers, the mediator activity of the full-length protein and for DNA repair in vivo in the presence of methyl methanesulfonate. Our studies suggested that phenylalanine-349 and tyrosine-409 are key residues in the C-terminal half of Rad52 and probably play an important role in the mediator activity. SN - 1362-4962 UR - https://www.unboundmedicine.com/medline/citation/24163251/Functional_analyses_of_the_C_terminal_half_of_the_Saccharomyces_cerevisiae_Rad52_protein_ L2 - https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gkt986 DB - PRIME DP - Unbound Medicine ER -