Variants of the human RAD52 gene confer defects in ionizing radiation resistance and homologous recombination repair in budding yeast
Authors:Alissa D. Clear1,2,3, Glenn M. Manthey1,2, Olivia Lewis4,5, Isabelle Y. Lopez4,6, Rossana Rico4,7, Shannon Owens8,9, M. Cristina Negritto10, Elise W. Wolf10,11, Jason Xu10,12, Nikola Kenjić13, J. Jefferson P. Perry13, Aaron W. Adamson14, Susan L. Neuhausen14, Adam M. Bailis1,2,15
doi: 10.15698/mic2020.10.732
Volume 7, pp. 270 to 285, published 20/07/2020.
1 Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, Duarte, CA, USA.
2 Irell & Manella Graduate School of Biological Sciences, Beckman Research Institute of City of Hope, Duarte, CA, USA.
3 bioStrategies Group, Chicago, IL, USA.
4 City of Hope – Duarte High School NIH Science Education Partnership Award Program, Duarte, CA, USA.
5 Barbara Bush Houston Literacy Foundation, Houston, TX, USA.
6 California State Polytechnic University at Pomona, Pomona, CA, USA.
7 Henry Samueli School of Engineering and Applied Sciences, University of California at Los Angeles, Los Angeles, CA, USA.
8 Eugene and Ruth Roberts Summer Student Academy, Beckman Research Institute of City of Hope, Duarte, CA, USA.
9 Department of Microbiology and Molecular Genetics, College of Biological Sciences, University of California at Davis, Davis, CA, USA.
10 Molecular Biology Program, Pomona College, Claremont, CA, USA.
11 Department of Microbiology and Immunology, University of California at San Francisco, San Francisco, CA, USA.
12 Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
13 Department of Biochemistry, University of California at Riverside, Riverside, CA, USA.
14 Department of Population Sciences, Beckman Research Institute of City of Hope, Duarte, CA, USA.
15 College of Health Professions, Thomas Jefferson University, Philadelphia, PA, USA.
Keywords:
HsRAD52 variants, tumorigenesis, DNA double strand breaks, homologous recombination repair, ionizing radiation, budding yeast
Corresponding Author(s):
Conflict of interest statement:
The authors declare that there are no conflicts of interest.
Please cite this article as:
Alissa D. Clear, Glenn M. Manthey, Olivia Lewis, Isabelle Y. Lopez, Rossana Rico, Shannon Owens, M. Cristina Negritto, Elise W. Wolf, Jason Xu, Nikola Kenjić, J. Jefferson P. Perry, Aaron W. Adamson, Susan L. Neuhausen, Adam M. Bailis (2020). Variants of the human RAD52 gene confer defects in ionizing radia-tion resistance and homologous recombination repair in budding yeast. Microbial Cell 7(10): 270-285. doi: 10.15698/mic2020.10.732
© 2020 Clear et al. This is an open-access article released under the terms of the Creative Commons Attribution (CC BY) license, which allows the unrestricted use, distribution, and reproduc-tion in any medium, provided the original author and source are acknowledged.
Abstract:
RAD52 is a structurally and functionally conserved component of the DNA double-strand break (DSB) repair apparatus from budding yeast to humans. We recently showed that expressing the human gene, HsRAD52 in rad52 mutant budding yeast cells can suppress both their ionizing radiation (IR) sensitivity and homologous recombination repair (HRR) defects. Intriguingly, we observed that HsRAD52 supports DSB repair by a mechanism of HRR that conserves genome structure and is independent of the canonical HR machinery. In this study we report that naturally occurring variants of HsRAD52, one of which suppresses the pathogenicity of BRCA2 mutations, were unable to suppress the IR sensitivity and HRR defects of rad52 mutant yeast cells, but fully suppressed a defect in DSB repair by single-strand annealing (SSA). This failure to suppress both IR sensitivity and the HRR defect correlated with an inability of HsRAD52 protein to associate with and drive an interaction between genomic sequences during DSB repair by HRR. These results suggest that HsRAD52 supports multiple, distinct DSB repair apparatuses in budding yeast cells and help further define its mechanism of action in HRR. They also imply that disruption of HsRAD52-dependent HRR in BRCA2-defective human cells may contribute to protection against tumorigenesis and provide a target for killing BRCA2-defective cancers.