Abstract
DNA double-strand break repair allows cells to survive both exogenous and endogenous insults to the genome. In yeast, the recombinases Rad51 and Rad52 are central to multiple forms of homology-dependent repair. Classically, Rad51 and Rad52 are thought to act cooperatively, with formation of the functional Rad51 nucleofilament facilitated by the mediator function of Rad52. Several studies have now identified functions for the interaction between Rad51 and Rad52 that are independent of the mediator function of Rad52 and affect a seemingly diverse array of functions in de novo telomere addition, global chromosome mobility following DNA damage, Rad51 nucleofilament stability, checkpoint adaptation, and microhomology-mediated chromosome rearrangements. Here, we review these functions with an emphasis on our recent discovery that the Rad51–Rad52 interaction influences the probability of de novo telomere addition at sites preferentially targeted by telomerase following a double-strand break (DSB). We present data addressing the prevalence of sites within the yeast genome that are capable of stimulating de novo telomere addition following a DSB and speculate about the potential role such sites may play in genome stability.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data Availability
Data summarized in Fig. 1 are provided in supplementary Table S1.
References
Adzuma K, Ogawa T, Ogawa H (1984) Primary structure of the RAD52 gene in Saccharomyces cerevisiae. Mol Cell Biol 4:2735–2744. https://doi.org/10.1128/mcb.4.12.2735
Antúnez de Mayolo A, Lisby M, Erdeniz N et al (2006) Multiple start codons and phosphorylation result in discrete Rad52 protein species. Nucleic Acids Res 34:2587–2597. https://doi.org/10.1093/nar/gkl280
Aylon Y, Kupiec M (2004) DSB repair: the yeast paradigm. DNA Repair (Amst) 3:797–815. https://doi.org/10.1016/J.DNAREP.2004.04.013
Bhargava R, Onyango DO, Stark JM (2016) Regulation of single-strand annealing and its role in genome maintenance. Trends Genet 32:566–575. https://doi.org/10.1016/j.tig.2016.06.007
Bordelet H, Dubrana K (2019) Keep moving and stay in a good shape to find your homologous recombination partner. Curr Genet 65:29–39
Churikov D, Géli V (2017) De novo telomere addition at chromosome breaks: dangerous liaisons. J Cell Biol 216:2243–2245. https://doi.org/10.1083/jcb.201705156
Coutelier H, Xu Z (2019) Adaptation in replicative senescence: a risky business. Curr Genet 65:711–716
Elango R, Sheng Z, Jackson J et al (2017) Break-induced replication promotes formation of lethal joint molecules dissolved by Srs2. Nat Commun 8:1–13. https://doi.org/10.1038/s41467-017-01987-2
Epum EA, Mohan MJ, Ruppe NP, Friedman KL (2020) Interaction of yeast Rad51 and Rad52 relieves Rad52-mediated inhibition of de novo telomere addition. PLoS Genet 16:e1008608. https://doi.org/10.1371/journal.pgen.1008608
Evans SK, Lundblad V (1999) Est1 and Cdc13 as comediators of telomerase access. Science 286:117–120. https://doi.org/10.1126/science.286.5437.117
Firmenich AA, Elias-Arnanz M, Berg P (1995) A novel allele of Saccharomyces cerevisiae RFA1 that is deficient in recombination and repair and suppressible by RAD52. Mol Cell Biol 15:1620–1631. https://doi.org/10.1128/mcb.15.3.1620
Fishman-Lobell J, Rudin N, Haber JE (1992) Two alternative pathways of double-strand break repair that are kinetically separable and independently modulated. Mol Cell Biol 12:1292–1303. https://doi.org/10.1128/mcb.12.3.1292
Fung CW, Fortin GS, Peterson SE, Symington LS (2006) The rad51-K191R ATPase-defective mutant Is impaired for presynaptic filament formation. Mol Cell Biol 26:9544–9554. https://doi.org/10.1128/mcb.00599-06
Gibb B, Ye LF, Gergoudis SC et al (2014a) Concentration-dependent exchange of replication protein A on single-stranded DNA revealed by single-molecule imaging. PLoS ONE 9:e87922. https://doi.org/10.1371/journal.pone.0087922
Gibb B, Ye LF, Kwon Y et al (2014b) Protein dynamics during presynaptic complex assembly on individual ssDNA molecules. Nat Struct Mol Biol 21:893. https://doi.org/10.1038/NSMB.2886
Hays SL, Firmenich AA, Berg P (1995) Complex formation in yeast double-strand break repair: participation of Rad51, Rad52, Rad55, and Rad57 proteins. Proc Natl Acad Sci U S A 92:6925–6929. https://doi.org/10.1073/pnas.92.15.6925
Hays SL, Firmenich AA, Massey P et al (1998) Studies of the interaction between Rad52 protein and the yeast single-stranded DNA binding protein RPA. Mol Cell Biol 18:4400–4406. https://doi.org/10.1128/mcb.18.7.4400
Jasin M, Rothstein R (2013) Repair of strand breaks by homologous recombination. Cold Spring Harb Perspect Biol 5:a012740. https://doi.org/10.1101/cshperspect.a012740
Kramara J, Osia B, Malkova A (2018) Break-induced replication: the where, the why, and the how. Trends Genet 34:518–531. https://doi.org/10.1016/j.tig.2018.04.002
Kramer KM, Haber JE (1993) New telomeres in yeast are initiated with a highly selected subset of TG1-3 repeats. Genes Dev 7:2345–2356. https://doi.org/10.1101/gad.7.12a.2345
Krejci L, Damborsky J, Thomsen B et al (2001) Molecular dissection of interactions between Rad51 and members of the recombination-repair group. Mol Cell Biol 21:966–976. https://doi.org/10.1128/MCB.21.3.966-976.2001
Krejci L, Song B, Bussen W et al (2002) Interaction with Rad51 is indispensable for recombination mediator function of Rad52. J Biol Chem 277:40132–40141. https://doi.org/10.1074/jbc.M206511200
Krogh BO, Symington LS (2004) Recombination proteins in yeast. Annu Rev Genet 38:233–271. https://doi.org/10.1146/annurev.genet.38.072902.091500
Lee SE, Moore JK, Holmes A et al (1998) Saccharomyces Ku70, Mre11/Rad50, and RPA proteins regulate adaptation to G2/M arrest after DNA damage. Cell 94:399–409. https://doi.org/10.1016/S0092-8674(00)81482-8
Lee SE, Pellicioli A, Malkova A et al (2001) The Saccharomyces recombination protein Tid1p is required for adaptation from G2/M arrest induced by a double-strand break. Curr Biol 11:1053–1057. https://doi.org/10.1016/S0960-9822(01)00296-2
Lee SE, Pellicioli A, Vaze MB et al (2003) Yeast Rad52 and Rad51 recombination proteins define a second pathway of DNA damage assessment in response to a single double-strand break. Mol Cell Biol 23:8913–8923. https://doi.org/10.1128/mcb.23.23.8913-8923.2003
Ma E, Dupaigne P, Maloisel L et al (2018) Rad52-Rad51 association is essential to protect Rad51 filaments against Srs2, but facultative for filament formation. Elife. https://doi.org/10.7554/eLife.32744
Mangahas JL, Alexander MK, Sandell LL, Zakian VA (2001) Repair of chromosome ends after telomere loss in Saccharomyces. Mol Biol Cell 12:4078–4089. https://doi.org/10.1091/mbc.12.12.4078
Mehta A, Haber JE (2014) Sources of DNA double-strand breaks and models of recombinational DNA repair. Cold Spring Harb Perspect Biol 6:a016428. https://doi.org/10.1101/cshperspect.a016428
Miné-Hattab J, Rothstein R (2013) DNA in motion during double-strand break repair. Trends Cell Biol 23:529–536
Myung K, Chen C, Kolodner RD (2001) Multiple pathways cooperate in the suppression of genome instability in Saccharomyces cerevisiae. Nature 411:1073–1076. https://doi.org/10.1038/35082608
Nugent CI, Hughes TR, Lue NF, Lundblad V (1996) Cdc13p: a single-strand telomeric DNA-binding protein with a dual role in yeast telomere maintenance. Science 274:249–252. https://doi.org/10.1126/science.274.5285.249
Obodo UC, Epum EA, Platts MH et al (2016) Endogenous hot spots of de novo telomere addition in the yeast genome contain proximal enhancers that bind Cdc13. Mol Cell Biol 36:1750–1763. https://doi.org/10.1128/mcb.00095-16
Ouenzar F, Lalonde M, Laprade H et al (2017) Cell cycle-dependent spatial segregation of telomerase from sites of DNA damage. J Cell Biol 216:2355–2371. https://doi.org/10.1083/jcb.201610071
Oza P, Jaspersen SL, Miele A et al (2009) Mechanisms that regulate localization of a DNA double-strand break to the nuclear periphery. Genes Dev 23:912–927. https://doi.org/10.1101/gad.1782209
Pennaneach V, Putnam CD, Kolodner RD (2006) Chromosome healing by de novo telomere addition in Saccharomyces cerevisiae. Mol Microbiol 59:1357–1368. https://doi.org/10.1111/j.1365-2958.2006.05026.x
Piazza A, Heyer WD (2019) Moving forward one step back at a time: reversibility during homologous recombination. Curr Genet 65:1333–1340
Putnam CD, Pennaneach V, Kolodner RD (2004) Chromosome healing through terminal deletions generated by de novo telomere additions in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A 101:13262–13267. https://doi.org/10.1073/pnas.0405443101
Rice C, Skordalakes E (2016) Structure and function of the telomeric CST complex. Comput Struct Biotechnol J 14:161–167
Sandell LL, Zakian VA (1993) Loss of a yeast telomere: arrest, recovery, and chromosome loss. Cell 75:729–739
Seong C, Colavito S, Kwon Y et al (2009) Regulation of Rad51 recombinase presynaptic filament assembly via interactions with the Rad52 mediator and the Srs2 anti-recombinase. J Biol Chem 284:24363–24371. https://doi.org/10.1074/jbc.M109.032953
Smith MJ, Bryant EE, Rothstein R (2018) Increased chromosomal mobility after DNA damage is controlled by interactions between the recombination machinery and the checkpoint. Genes Dev 32:1242–1251. https://doi.org/10.1101/gad.317966.118
Stellwagen AE, Haimberger ZW, Veatch JR, Gottschling DE (2003) Ku interacts with telomerase RNA to promote telomere addition at native and broken chromosome ends. Genes Dev 17:2384–2395. https://doi.org/10.1101/gad.1125903
Sugawara N, Wang X, Haber JE (2003) In vivo roles of Rad52, Rad54, and Rad55 proteins in Rad51-mediated recombination. Mol Cell 12:209–219
Sugiyama T, Kantake N (2009) Dynamic regulatory interactions of Rad51, Rad52, and Replication Protein-A in recombination intermediates. J Mol Biol 390:45–55. https://doi.org/10.1016/J.JMB.2009.05.009
Sung P, Stratton SA (1996) Yeast Rad51 recombinase mediates polar DNA strand exchange in the absence of ATP hydrolysis. J Biol Chem 271:27983–27986. https://doi.org/10.1074/jbc.271.45.27983
Toczyski DP, Galgoczy DJ, Hartwell LH (1997) CDC5 and CKII control adaptation to the yeast DNA damage checkpoint. Cell 90:1097–1106. https://doi.org/10.1016/S0092-8674(00)80375-X
Umezu K, Sugawara N, Chen C et al (1998) Genetic analysis of yeast RPA1 reveals its multiple functions in DNA metabolism. Genetics 148:989–1005
Vasianovich Y, Altmannova V, Kotenko O et al (2017) Unloading of homologous recombination factors is required for restoring double‐stranded DNA at damage repair loci. EMBO J 36:213–231. https://doi.org/10.15252/embj.201694628
Vaze MB, Pellicioli A, Lee SE et al (2002) Recovery from checkpoint-mediated arrest after repair of a double-strand break requires Srs2 helicase. Mol Cell 10:373–385. https://doi.org/10.1016/S1097-2765(02)00593-2
Waterman DP, Haber JE, Smolka MB (2020) Checkpoint responses to DNA double-strand breaks. Annu Rev Biochem. https://doi.org/10.1146/annurev-biochem-011520-104722
Wu Y, Kantake N, Sugiyama T, Kowalczykowski SC (2008) Rad51 protein controls Rad52-mediated DNA annealing. J Biol Chem 283:14883–14892. https://doi.org/10.1074/jbc.M801097200
Zhang W, Durocher D (2010) De novo telomere formation is suppressed by the Mec1-dependent inhibition of Cdc13 accumulation at DNA breaks. Genes Dev 24:502–515. https://doi.org/10.1101/gad.1869110
Zimmer C, Fabre E (2019) Chromatin mobility upon DNA damage: state of the art and remaining questions. Curr Genet. https://doi.org/10.1007/s00294-018-0852-6
Acknowledgements
We thank Sara Conwell and Blake Conwell for technical assistance. Many thanks to Dr. James Haber for gifts of strains and plasmids and for insightful comments on the manuscript.
Funding
This work is supported by National Institutes of Health award R01GM123292 to KLF.
Author information
Authors and Affiliations
Contributions
KLF wrote the manuscript; KN and EAE contributed to the writing and ideas expressed in the manuscript; KN generated the data.
Corresponding author
Ethics declarations
Conflicts of interest
The authors have declared that no competing interests exist.
Additional information
Communicated by M. Kupiec.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Ngo, K., Epum, E.A. & Friedman, K.L. Emerging non-canonical roles for the Rad51–Rad52 interaction in response to double-strand breaks in yeast. Curr Genet 66, 917–926 (2020). https://doi.org/10.1007/s00294-020-01081-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00294-020-01081-z