Skip to main content
Log in

Partial purification of an activity from human cells that promotes homologous pairing and the formation of heteroduplex DNA in the presence of ATP

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

An activity that can promote homologous pairing and strand transfer between suitable DNA substrates has been partially purified from human skin fibroblasts and from Hela cells. The strand transfer reaction was investigated with DNA substrates consisting of single-stranded circular and duplex linear phage DNA. It requires ATP, and under optimal conditions yields heteroduplex molecules containing one strand from each parental DNA substrate. The reactions appears to be of the same general nature as those mediated by RecA proteins of Escherichia coli and the Rec1 protein of Ustilago maydis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ayares D, Spencer J, Schwartz F, Morse B, Kucherlapati R (1985) Homologous recombination between autonomously replicating plasmids in mammalian cells. Genetics 111:375–388

    Google Scholar 

  • Cassuto E, West SC, Mursalim J, Conlon S, Howard-Flanders P (1980) Initiation of genetic recombination: Homologous pairing between duplex DNA molecules in the presence of RecA protein. Proc Natl Acad Sci USA 77:3962–3966

    Google Scholar 

  • Cox MM, Lehman IR (1981) RecA protein of Escherichia coli promotes branch migration, a kinetically distinct phase of DNA strand exchange. Proc Natl Acad Sci USA 78:3433–3437

    Google Scholar 

  • Cunningham RP, DasGupta C, Shibata T, Radding CM (1980) Homologous pairing in genetic recombination: recA protein makes joint molecules of gapped circular DNA and closed circular DNA. Cell 20:223–235

    Google Scholar 

  • De Saint Vincent BR, Wahl G (1983) Homologous recombination in mammalian cells mediates formation of functional gene from two overlapping fragments. Proc Natl Acad Sci USA 80:2002–2006

    Google Scholar 

  • Folger KR, Wong EA, Wahl G, Capecchi MR (1982) Patterns of integration of DNA microinjected into cultured mammalian cells: Evidence for homologous recombination between injected plasmid DNA molecules. Mol Cell Biol 2:1372–1387

    Google Scholar 

  • Hamelin C, Alain R, Houde D, Berthiaume L (1983) Mammalian cells contain a DNA-recombining activity that can be dissociated from topoisomerase activity. Biosci Rep 2:527–533

    Google Scholar 

  • Hsieh P, Meyn MS, Camerini-Otero D (1986) Partial purification and characterization of a recombinase from human cells. Cell 44:885–894

    Google Scholar 

  • Kahn R, Cunningham RP, DasGupta C, Radding CM (1981) Polarity of heteroduplex formation promoted by Escherichia coli recA protein. Proc Natl Acad Sci USA 78:4786–4790

    Google Scholar 

  • Kenne K, Lindquist S (1984) A DNA-recombinogenic activity in human cells. Nucleic Acids Res 12:3057–3068

    Google Scholar 

  • Kmiec E, Holloman WK (1982) Homologous pairing of DNA molecules promoted by a protein from Ustilago. Cell 29:367–374

    Google Scholar 

  • Kmiec E, Holloman WK (1983) Heteroduplex formation and polarity during strand transfer promoted by Ustilago Rec1 protein. Cell 33:857–864

    Google Scholar 

  • Miller CL, Temin HM (1983) High efficiency ligation and recombination of DNA fragments by vertebrate cells. Science 220:606–609

    Google Scholar 

  • Orr-Weaver TL, Szostak JW, Rothstein RJ (1981) Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci USA 78:6354–6358

    Google Scholar 

  • Shibata T, Cunningham RP, DasGupta C, Radding CM (1979) Homologous pairing in genetic recombination: complexes of recA protein and DNA. Proc Natl Acad Sci USA 76:5100–5104

    Google Scholar 

  • Small J, Scangos G (1983) Recombination during gene transfer into mouse cells can restore the function of deleted genes. Science 219:174–176

    Google Scholar 

  • Song K-Y, Chekuri L, Rauth S, Ehrlich S, Kucherlapati R (1985) Effect of double-strand breaks on homologous recombination in mammalian cells and extracts. Mol Cell Biol 5:3331–3336

    Google Scholar 

  • Upcroft P, Carter B, Kidson C (1980) Analysis of recombination in mammalian cells using SV40 genome segments having homologous overlapping termini. Nucleic Acids Res 8:2725–2735

    Google Scholar 

  • Wake CT, Wilson JH (1979) Simian virus 40 recombinants are produced at high frequency during infection with genetically mixed oligomeric DNA. Proc Natl Acad Sci USA 76:2876–2880

    Google Scholar 

  • West SC, Cassuto E, Howard-Flanders P (1981) RecA protein promotes homologous pairing and strand exchange reactions between duplex DNA molecules. Proc Natl Acad Sci USA 78:2100–2104

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by G.R. Smith

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cassuto, E., Lightfoot, LA. & Howard-Flanders, P. Partial purification of an activity from human cells that promotes homologous pairing and the formation of heteroduplex DNA in the presence of ATP. Mole Gen Genet 208, 10–14 (1987). https://doi.org/10.1007/BF00330415

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330415

Key words

Navigation