DNA Strand Annealing is Promoted by the Yeast Rad52 Protein

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1996 Oct 1

PMID 8855248

Citations 248

Authors

U H Mortensen

C Bendixen

I Sunjevaric

R Rothstein

Affiliations

Soon will be listed here.

Abstract

The Saccharomyces cerevisiae RAD52 gene plays a pivotal role in genetic recombination. Here we demonstrate that yeast Rad52 is a DNA binding protein. To show that the interaction between Rad52 and DNA is direct and not mediated by other yeast proteins and to facilitate protein purification, a recombinant expression system was developed. The recombinant protein can bind both single- and double-stranded DNA and the addition of either Mg2+ or ATP does not enhance the binding of single-stranded DNA. Furthermore, a DNA binding domain was found in the evolutionary conserved N terminus of the protein. More importantly, we show that the protein stimulates DNA annealing even in the presence of a large excess of nonhomologous DNA. Rad52-promoted annealing follows second-order kinetics and the rate is 3500-fold faster than that of the spontaneous reaction. How this annealing activity relates to the genetic phenotype associated with rad52 mutant cells is discussed.

Citing Articles

Mechanism of single-stranded DNA annealing by RAD52-RPA complex.

Liang C, Greenhough L, Masino L, Maslen S, Bajrami I, Tuppi M Nature. 2024; 629(8012):697-703.

PMID: 38658755 PMC: 11096129. DOI: 10.1038/s41586-024-07347-7.

PCNA Unloading Is Crucial for the Bypass of DNA Lesions Using Homologous Recombination.

Arbel-Groissman M, Liefshitz B, Katz N, Kuryachiy M, Kupiec M Int J Mol Sci. 2024; 25(6).

PMID: 38542333 PMC: 10970473. DOI: 10.3390/ijms25063359.

Bridging the gap: R-loop mediated genomic instability and its implications in neurological diseases.

Westover K, Jin P, Yao B Epigenomics. 2024; .

PMID: 38530068 PMC: 11160457. DOI: 10.2217/epi-2023-0379.

Gross Chromosomal Rearrangement at Centromeres.

Xu R, Pan Z, Nakagawa T Biomolecules. 2024; 14(1).

PMID: 38254628 PMC: 10813616. DOI: 10.3390/biom14010028.

Yeast Rad52 is a homodecamer and possesses BRCA2-like bipartite Rad51 binding modes.

Deveryshetty J, Chadda R, Mattice J, Karunakaran S, Rau M, Basore K Nat Commun. 2023; 14(1):6215.

PMID: 37798272 PMC: 10556141. DOI: 10.1038/s41467-023-41993-1.

References

Pontius B, Berg P . Renaturation of complementary DNA strands mediated by purified mammalian heterogeneous nuclear ribonucleoprotein A1 protein: implications for a mechanism for rapid molecular assembly. Proc Natl Acad Sci U S A. 1990; 87(21):8403-7. PMC: 54964. DOI: 10.1073/pnas.87.21.8403. View

Johnson R, Symington L . Functional differences and interactions among the putative RecA homologs Rad51, Rad55, and Rad57. Mol Cell Biol. 1995; 15(9):4843-50. PMC: 230729. DOI: 10.1128/MCB.15.9.4843. View

Shinohara A, Ogawa H, Ogawa T . Rad51 protein involved in repair and recombination in S. cerevisiae is a RecA-like protein. Cell. 1992; 69(3):457-70. DOI: 10.1016/0092-8674(92)90447-k. View

Ogawa T, Yu X, Shinohara A, Egelman E . Similarity of the yeast RAD51 filament to the bacterial RecA filament. Science. 1993; 259(5103):1896-9. DOI: 10.1126/science.8456314. View

Connolly B, Parsons C, Benson F, Dunderdale H, Sharples G, Lloyd R . Resolution of Holliday junctions in vitro requires the Escherichia coli ruvC gene product. Proc Natl Acad Sci U S A. 1991; 88(14):6063-7. PMC: 52022. DOI: 10.1073/pnas.88.14.6063. View

Resnick M . Genetic control of radiation sensitivity in Saccharomyces cerevisiae. Genetics. 1969; 62(3):519-31. PMC: 1212294. DOI: 10.1093/genetics/62.3.519. View

Carlson M, Laurent B . The SNF/SWI family of global transcriptional activators. Curr Opin Cell Biol. 1994; 6(3):396-402. DOI: 10.1016/0955-0674(94)90032-9. View

Hays S, Firmenich A, Berg P . Complex formation in yeast double-strand break repair: participation of Rad51, Rad52, Rad55, and Rad57 proteins. Proc Natl Acad Sci U S A. 1995; 92(15):6925-9. PMC: 41443. DOI: 10.1073/pnas.92.15.6925. View

Resnick M, Nitiss J, Edwards C, Malone R . Meiosis can induce recombination in rad52 mutants of Saccharomyces cerevisiae. Genetics. 1986; 113(3):531-50. PMC: 1202854. DOI: 10.1093/genetics/113.3.531. View

10.

Thomas B, Rothstein R . Elevated recombination rates in transcriptionally active DNA. Cell. 1989; 56(4):619-30. DOI: 10.1016/0092-8674(89)90584-9. View

11.

Firmenich A, Berg P . A novel allele of Saccharomyces cerevisiae RFA1 that is deficient in recombination and repair and suppressible by RAD52. Mol Cell Biol. 1995; 15(3):1620-31. PMC: 230386. DOI: 10.1128/MCB.15.3.1620. View

12.

Ogawa T, Shinohara A, Nabetani A, Ikeya T, Yu X, Egelman E . RecA-like recombination proteins in eukaryotes: functions and structures of RAD51 genes. Cold Spring Harb Symp Quant Biol. 1993; 58:567-76. DOI: 10.1101/sqb.1993.058.01.063. View

13.

Friedberg E . Eukaryotic DNA repair: glimpses through the yeast Saccharomyces cerevisiae. Bioessays. 1991; 13(6):295-302. DOI: 10.1002/bies.950130607. View

14.

Game J . DNA double-strand breaks and the RAD50-RAD57 genes in Saccharomyces. Semin Cancer Biol. 1993; 4(2):73-83. View

15.

Ostermann K, Lorentz A, Schmidt H . The fission yeast rad22 gene, having a function in mating-type switching and repair of DNA damages, encodes a protein homolog to Rad52 of Saccharomyces cerevisiae. Nucleic Acids Res. 1993; 21(25):5940-4. PMC: 310478. DOI: 10.1093/nar/21.25.5940. View

16.

Malone R, Esposito R . The RAD52 gene is required for homothallic interconversion of mating types and spontaneous mitotic recombination in yeast. Proc Natl Acad Sci U S A. 1980; 77(1):503-7. PMC: 348300. DOI: 10.1073/pnas.77.1.503. View

17.

Sung P, Robberson D . DNA strand exchange mediated by a RAD51-ssDNA nucleoprotein filament with polarity opposite to that of RecA. Cell. 1995; 82(3):453-61. DOI: 10.1016/0092-8674(95)90434-4. View

18.

Prakash S, Prakash L, Burke W, Montelone B . Effects of the RAD52 Gene on Recombination in SACCHAROMYCES CEREVISIAE. Genetics. 1980; 94(1):31-50. PMC: 1214136. DOI: 10.1093/genetics/94.1.31. View

19.

Bezzubova O, Schmidt H, Ostermann K, Heyer W, Buerstedde J . Identification of a chicken RAD52 homologue suggests conservation of the RAD52 recombination pathway throughout the evolution of higher eukaryotes. Nucleic Acids Res. 1993; 21(25):5945-9. PMC: 310479. DOI: 10.1093/nar/21.25.5945. View

20.

Rattray A, Symington L . Use of a chromosomal inverted repeat to demonstrate that the RAD51 and RAD52 genes of Saccharomyces cerevisiae have different roles in mitotic recombination. Genetics. 1994; 138(3):587-95. PMC: 1206210. DOI: 10.1093/genetics/138.3.587. View