Plant Molecular Biology

, Volume 57, Issue 1, pp 1–11 | Cite as

DNA binding and pairing activity of OsDmc1, a recombinase from rice



A cloned cDNA corresponding to OsDMC1 from rice anther tissue was expressed in Escherichia coli. The OsDmc1 protein was largely present in the inclusion bodies of the cell lysatE., which was solubilized by 8.0 M urea containing buffeR., purified to homogeneity by Ni-CAM agarose column chromatography, followed by renaturation to its native state through stepwise dialysis against reduced concentrations of urea. The purified protein cross-reacted with anti-yeast Dmc1 antibodies. The binding efficiency observed with circular single-stranded DNA (ssDNA) was similar to that with circular double-stranded DNA (dsDNA). The binding to either DNA showed no ATP dependencE., but required 5–10 mM Mg2+ in the presence of ATP. Even though the protein binding to dsDNA was as efficient as it was to ssDNA, the former induced no DNA dependent ATPasE., whereas the binding to ssDNA stimulated a significant level of DNA dependent ATPase activity. OsDmc1–ssDNA complex, with its ATPase proficiency, also mediated renaturation of homologous complementary strands as well as assimilation of single strands into homologous supercoiled duplexes leading to D-loop formation. The D-loop formation was lowered by excess of OsDmc1 protein. This D-loop formation activity was promoted by non-hydrolyzable ATP analog, AMP-PNP and was not observed in absence of ATP or presence of ADP/ATP-γ-S. These properties reflected the classical hallmarks of a recombinase and represented the first biochemical characterization of a plant Dmc1 protein.


ATPasE. D-loop DNA binding OsDmc1 Recombinase. Renaturation 



γ-S: Adenosine 5′-O-(3-thio-triphosphate)




Bovine serum albumin




Nickel chelating affinity matrix








replicative form


Oryza sativa disrupted meiotic cDNA1


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bishop, D.K., Park, D., Xu, L., Kleckner, N. 1992DMC1: a meiosis-specific yeast homolog of E. coli recA required for recombination, synaptonemal complex formation, and cell cycle progressionCell69439456Google Scholar
  2. Couteau, F., Belzile, F., Horlow, C., Grandjean, O., Vezon, D., Doutriaux, M.P. 1999Random chromosome segregation without meiotic arrest in both male and female meiocytes of a dmc1 mutant of ArabidopsisPlant Cell1116231634Google Scholar
  3. Ding, Z., Wang, T., Chong, K., Bai, S. 2001Isolation and characterization of OsDMC1, the rice homologue of the yeast DMC1 gene essential for meiosisSex Plant Reprod13285288Google Scholar
  4. Doutriaux, M.P, Couteau, F., Bergounioux, C., White, C. 1998Isolation and characterization of the RAD51 and DMC1 homologues from Arabidopsis thalianaMol Gen Genet257283291Google Scholar
  5. Gupta, R.C., Golub, E., Bi, B., Radding, C.M. 2001The synaptic activity of HsDmc1, a human recombination protein specific to meiosisProc Natl Acad Sci USA9884338439Google Scholar
  6. Hong, E.L, Shinohara, A., Bishop, D.K. 2001Saccharomyces cerevisiae Dmc1 protein promotes renaturation of single-strand DNA (ssDNA) and assimilation of ssDNA into homologous super-coiled duplex DNAJ Biol Chem2764190641912Google Scholar
  7. Kagawa, W., Kurumizaka, H., Ikawa, S., Yokoyama, S., Shibata, T. 2001Homologous pairing promoted by the human Rad52 proteinJ Biol Chem2763520135208Google Scholar
  8. Kathiresan, A, Khush, G.S., Bennett, J. 2002Two rice DMC1 genes are differentially expressed during meiosis and during haploid and diploid mitosisSex. Plant Reprod14257267Google Scholar
  9. Kinebuchi, T., Kagawa, W., Enomoto, R., Tanaka, K., Miyagawa, K., Shibata, T., Kurumizaka, H., Yokoyama, S. 2004Structural basis for octameric ring formation and DNA interaction of the human homologous-pairing protein Dmc1Mol. Cell14363374Google Scholar
  10. Kobayashi, T., Kobayashi, E., Sato, S., Hotta, Y, Miyajima, N, Tanaka, A., Tabata, S. 1994Characterization of cDNAs induced in meiotic prophase in lily microsporocytesDNA Res11526Google Scholar
  11. Kowalczykowski, S.C., Dixon, D.A, Eggleston, A.K., Lauder, S.D., Rehrauer, W.M. 1994Biochemistry of homologous recombination in Escherichia coliMicrobiol Rev58401465PubMedGoogle Scholar
  12. Laemmli, U.K. 1970Cleavage of structural proteins during the assembly of the head of bacteriophage. T4Nature le-parent227680685Google Scholar
  13. Li, Z., Golub, E.I, Gupta, R., Radding, C.M. 1997Recombination activities of HsDmc1 protein, the meiotic human homolog of RecA proteinProc Natl Acad Sci USA941122111226Google Scholar
  14. Masson, J.Y, Davies, A.A, Hajibagheri, N, VanDyck, E., Benson, F.E., Stasiak, A.Z., Stasiak, A., West, S.C. 1999The meiosis specific recombinase hDmc1 forms ring structures and interacts with hRad51EMBO J1865526560Google Scholar
  15. Masson, J.Y., West, S.C. 2001The Rad51 and Dmc1 recombinases: a non-identical twin relationshipTrends Biochem Sci26131136Google Scholar
  16. Metkar, S.S., Sainis, J.K., Mahajan, S.K. 2004Cloning and characterization of the DMC1 genes in Oryza sativaCurr Sci87353357Google Scholar
  17. Nara, T., Hamada, F., Namekawa, S., Sakaguchi, K. 2001Strand exchange reaction in vitro and DNA-dependent ATPase activity of recombinant LIM15/DMC1 and RAD51 proteins from Coprinus cinereusBiochem Biophys Res Commun2859297Google Scholar
  18. Nara, T., Yamamoto, T., Sakaguchi, K. 2000Characterization of interaction of C- and N-terminal domains in LIM15/DMC1 and RAD51 from a basidiomyceteS., Coprinus cinereusBiochem Biophys Res Commun27597102Google Scholar
  19. Passy, S.I, Yu, X, Li, Z., Radding, C.M, Masson, J.Y, West, S.C., Egelman, E.H. 1999Human Dmc1 protein binds DNA as octameric ringProc Natl Acad Sci USA961068410688Google Scholar
  20. Sato, S., Hotta, Y., Tabata, S. 1995Structural analysis of a recA-like gene in the genome of Arabidopsis thalianaDNA Res28993Google Scholar
  21. Sehorn, M.G, Sigurdsson, S., Bussen, W., Unger, V.M., Sung, P. 2004Human meiotic recombinase Dmc1 promotes ATP-dependent homologous DNA strand exchangeNature429433437Google Scholar
  22. Shimazu, J, Matsukura, C., Senda, M, Ishikawa, R., Akada, S., Harada, T., Tabata, S., Niizeki, M. 2001Characterization of a DMC1 homologuE., RiLIM15, in meiotic panicleS., mitotic cultured cells and mature leaves of rice (Oryza sativa L)Theor Appl Genet10211591163Google Scholar
  23. Symington, L.S. 2002Role of RAD52 epistasis group genes in homologous recombination and double strand break repairMicrobiol. Mol Biol Rev66630670Google Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  1. 1.Molecular Biology DivisionBhabha Atomic Research CenterMumbaiIndia
  2. 2.Department of Biological SciencesTata Institute of Fundamental ResearchMumbaiIndia

Personalised recommendations