Abstract
Rad51/RadA paralogs found in eukaryotes and euryarchaea play important roles during recombination and repair, and mutations in one of the human Rad51 paralogs, Rad51C, are associated with breast and ovarian cancers. The hyperthermophilic crenarchaeon Sulfolobus tokodaii encodes four putative RadA paralogs and studies on these proteins may assist in understanding the functions of human Rad51 paralogs. Here, we report the biochemical characterization of stRadC2, a S. tokodaii RadA paralog. Pull-down assays revealed that the protein was able to interact with the recombinase, RadA, and the Holliday junction endonuclease, Hjc. stRadC2 inhibited the strand exchange activity of RadA and facilitated Hjc-mediated Holliday junction DNA cleavage in vitro. RT-PCR analysis revealed that stRadC2 transcription was immediately reduced after UV irradiation, but was restored to normal levels at the late stages of DNA repair. Our results suggest that stRadC2 may act as an antirecombination factor in DNA recombinational repair in S. tokodaii.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
McCready S J, Osman F, Yasui A. Repair of UV damage in the fission yeast Schizosaccharomyces pombe. Mutat Res, 2000, 451: 197–210
Sung P. Yeast Rad55 and Rad57 proteins form a heterodimer that functions with replication protein A to promote DNA strand exchange by Rad51 recombinase. Genes Dev, 1997, 11: 1111–1121
Mozlin A M, Fung C W, Symington L S. Role of the Saccharomyces cerevisiae Rad51 paralogs in sister chromatid recombination. Genetics, 2008, 178: 113–126
Kawabata M, Kawabata T, Nishibori M. Role of recA/RAD51 family proteins in mammals. Acta Med Okayama, 2005, 59: 1–9
Thacker J. A surfeit of RAD 51-like genes? Trends Genet, 1999, 15: 166–168
Miller K A, Sawicka D, Barsky D. Domain mapping of the Rad51 paralog protein complexes. Nucleic Acids Res, 2004, 32: 169–178
Takata M, Sasaki M S, Tachiiri S, et al. Chromosome instability and defective recombinational repair in knockout mutants of the five Rad51 paralogs. Mol Cell Biol, 2001, 21: 2858–2866
Kelman Z, White M F. Archaeal DNA replication and repair. Curr Opin Microbiol, 2005, 8: 669–676
Haldenby S, White M F, Allers T. RecA family proteins in archaea: RadA and its cousins. Biochem Soc Trans, 2009, 37: 102–107
Komori K, Miyata T, DiRuggiero J, et al. Both RadA and RadB are involved in homologous recombination in Pyrococcus furiosus. J Biol Chem, 2000, 275: 33782–33790
Guy C P, Haldenby S, Brindley A, et al. Interactions of RadB, a DNA repair protein in archaea, with DNA and ATP. J Mol Biol, 2006, 358: 46–56
Sheng D, Zhu S, Wei T, et al. The in vitro activity of a Rad55 homologue from Sulfolobus tokodaii, a candidate mediator in RadA-catalyzed homologous recombination. Extremophiles, 2008, 12: 147–157
McRobbie A M, Carter L G, Kerou M, et al. Structural and functional characterisation of a conserved archaeal RadA paralog with antirecombinase activity. J Mol Biol, 2009, 389: 661–673
Abella M, Rodríguez S, Paytubi S, et al. The Sulfolobus solfataricus radA paralogue sso0777 is DNA damage inducible and positively regulated by the Sta1 protein. Nucleic Acids Res, 2007, 35: 6788–6797
Brock T D, Brock K M, Belly R T, et al. Sulfolobus: A new genus of sulfur-oxidizing bacteria living at low pH and high temperature. Arch Mikrobiol, 1972, 84: 54–68
Ausubel F M, Brent R, Kingston R E. Short Protocols in Molecular Biology. 3rd ed. New York: Cold Spring Harbor Laboratory Press, 1995
Zhang S, Wei T, Hou G, et al. Archaeal DNA helicase HerA interacts with Mre11 homologue and unwinds blunt-ended double-stranded DNA and recombination intermediates. DNA Repair (Amst), 2008, 7: 380–391
Komori K, Sakae S, Fujikane R, et al. Biochemical characterization of the Hjc Holliday junction resolvase of Pyrococcus furiosus. Nucleic Acids Res, 2000, 28: 4544–4551
Kawarabayasi Y, Hino Y, Horikawa H, et al. Complete genome sequence of an aerobic thermoacidophilic crenarchaeon, Sulfolobus tokodaii strain7. DNA Res, 2001, 8: 123–140
Kvaratskhelia M, White M F. Two Holliday junction resolving enzymes in Sulfolobus solfataricus. J Mol Biol, 2000, 297: 923–932
Sheng D, Zhu S, Li M, et al. Cloning, expression and radiation inducibility of RadA from the hyperthermophilic archaeon Sulfolobus tokodaii. Wei Sheng Wu Xue Bao, 2008, 48: 317–22
Klein H L. The consequences of Rad51 overexpression for normal and tumor cells. DNA Repair (Amst), 2008, 7: 686–693
Venkatesh R, Ganesh N, Guhan N, et al. RecX protein abrogates ATP hydrolysis and strand exchange promoted by RecA: insights into negative regulation of homologous recombination. Proc Natl Acad Sci USA, 2002, 99: 12091–12096
Lusetti S L, Drees J C, Stohl E A, et al. The DinI and RecX proteins are competing modulators of RecA function. J Biol Chem, 2004, 279: 55073–55079
Yoon D, Wang Y, Stapleford K, et al. P53 inhibits strand exchange and replication fork regression promoted by human Rad51. J Mol Biol, 2004, 336: 639–654
Houtgraaf J H, Versmissen J, van der Giessen W J. A concise review of DNA damage checkpoints and repair in mammalian cells. Cardiovasc Revasc Med, 2006, 7: 165–172
Author information
Authors and Affiliations
Corresponding author
Additional information
Contributed equally to this work
This article is published with open access at Springerlink.com
Rights and permissions
Open Access This article is distributed under the terms of the Creative Commons Attribution 2.0 International License (https://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
About this article
Cite this article
Wang, L., Sheng, D., Han, W. et al. Sulfolobus tokodaii RadA paralog, stRadC2, is involved in DNA recombination via interaction with RadA and Hjc. Sci. China Life Sci. 55, 261–267 (2012). https://doi.org/10.1007/s11427-012-4292-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11427-012-4292-0