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DNA cruciforms facilitate in vitro strand transfer on nucleosomal templates

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Abstract

A single, phased nucleosome assembled on a 240 by DNA duplex molecule blocked Escherichia coli RecA protein-promoted strand transfer of the complementary strand of the duplex onto a homologous single-stranded circle. However, when a four-armed cruciform structure was coupled to either end of the duplex the barrier to strand transfer was overcome and joint molecules were efficiently formed. Micrococcal nuclease digestion indicated that the nucleosome was dissociated by the juxtaposition of the cruciform. We interpret these results to mean that cruciform structures can act over a distance to destabilize adjacent nucleosomes and suggest that, as a consequence, the chromatin structure surrounding a crossed strand recombination intermediate might be disrupted, enabling other recombination events to initiate or the process of branch migration to proceed.

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References

  • Brown DD (1984) The role of stable complexes that repress and activate eucaryotic genes. Cell 37:359–365

    Google Scholar 

  • Chow SA, Honigberg SM, Bainton RJ, Radding CM (1986) Patterns of nuclease protection during strand exchange. J Biol Chem 261:6961–6971

    Google Scholar 

  • Clark DJ, Felsenfeld G (1992) A nucleosome core is transferred out of the path of a transcribing polymerase. Cell 71:11–22

    Google Scholar 

  • Cox MM, Lehman IR (1987) Enzymes of general recombination. Annu Rev Biochem 56:223–262

    Google Scholar 

  • Germond RP, Bellard M, Oudet P, Chambon P (1976) Stability of nucleosomes in native and reconstituted chromatins. Nucleic Acids Res 3:3173–3192

    Google Scholar 

  • Gonda DK, Radding CM (1983) By searching processively RecA protein pairs DNA molecules that share a limited stretch of homology. Cell 34:647–654

    Google Scholar 

  • Gonda DK, Radding CM (1986) The mechanism of the search for homology promoted by recA protein. Facilitated diffusion within nucleoprotein networks. J Biol Chem 261:13087–13096

    Google Scholar 

  • van Holde KE (1989) Chromatin. Springer-Verlag Publishers, New York

    Google Scholar 

  • Holliday R (1964) A mechanism for gene conversion in fungi. Genet Res 5:282–304

    Google Scholar 

  • Honigberg SM, Gonda DK, Radding CM (1985) The pairing activity of stable nucleoprotein filaments made from recA protein, single-stranded DNA, and adenosine 5′-(gammathio)triphosphate. J Biol Chem 260:11845–11851

    Google Scholar 

  • Kahn R, Radding CM (1984) Separation of the presynaptic and synaptic phases of homologous pairing promoted by recA protein. J Biol Chem 259:7495–7503

    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 

  • Kawasaki I, Sugano S, Ikeda H (1989) Calf thymus histone H1 is a recombinase that catalyzes ATP-independent DNA strand transfer. Proc Natl Acad Sci USA 86:5281–5285

    Google Scholar 

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

    Google Scholar 

  • Kmiec EB, Holloman WK ATP-dependent DNA renaturation and DNA-dependent ATPase reactions catalyzed by the Ustilago maydis homologous pairing protein. Eur J Biochem 219:865–875

  • Kmiec EB, Sekiguchi JM, Cole AD (1989) Studies on the ATP requirements of in vitro chromatin assembly. Biochem Cell Biol 67:443–454

    Google Scholar 

  • Kotani H, Kmiec EB Transcription activates RecA-promoted homologous pairing of nucleosomal DNA. Mol Cell Biol 3:1949–1955

  • Lorch Y, LaPoint JW, Kornberg RD (1987) Nucleosomes inhibit the initiation of transcription but allow chain elongation with the displacement of histones. Cell 49:203–210

    Google Scholar 

  • McCarthy JG, Sander M, Lowenhaupt K, Rich A (1988) Sensitive homologous recombination strand-transfer assay: partial purification of a Drosophila melanogaster enzyme and detection of sequence effects on the strand-transfer activity of recA protein. Proc Natl Acad Sci USA 85:5854–5858

    Google Scholar 

  • Morse RH (1988) Nucleosomes inhibit both transcriptional initiation and elongation by RNA polymerase III in vitro. EMBO J 8:2343–2351

    Google Scholar 

  • O'Neill TE, Roberge M, Bradbury EM (1992) Nucleosome arrays inhibit both initiation and elongation of transcripts by bacteriophage T7 RNA polymerase. J Mol Biol 223:67–78

    Google Scholar 

  • Radding CM (1988) Homologous pairing and strand exchange promoted by Escherichia coli recA protein. In: Kucherlapati R, Smith GR (eds) Genetic Recombination. American Sociaty for Microbiology, Washington, D.C., pp 193–229

    Google Scholar 

  • Ramdas J, Mythili E, Muniyappa K (1991) Nucleosomes on linear duplex DNA allow homologous pairing but prevent strand exchange promoted by recA protein. Proc Natl Acad Sci USA 88:1344–1348

    Google Scholar 

  • Razvi F, Garguilo G, Worcel A (1983) A simple procedure for parallel sequencing of both strands of 5′ labeled DNA. Gene 23:175–183

    Google Scholar 

  • Roca AI, Cox MM (1990) The recA protein: structure and function. Crit Rev Biochem Mol Biol 25:415–456

    Google Scholar 

  • Simon RH, Felsenfeld G (1979) A new procedure for purifying histone pairs H2A + H2B and H3 + H4 from chromatin using hydroxylapatite. Nucleic Acids Res 6:689–714

    Google Scholar 

  • Svaren J, Inagami S, Lovegren E, Chalkley R (1987) DNA denatures upon drying after ethanol precipitation. Nucleic Acids Res 15:8739–8753

    Google Scholar 

  • Wahls WP, Wallace LJ, Morre PD (1990) The Z-DNA motif d(TG) 30 promotes reception of information during gene conversion events while stimulating homologous recombination in human cells in culture. Mol Cell Biol 10:785–793

    Google Scholar 

  • Wang JNC, Hogan M, Austin RH (1982) DNA motions in the nucleosome core particle. Proc Natl Acad Sci USA 79:5896–5900

    Google Scholar 

  • West SC, Cassuto E, Howard-Flanders P (1981) Heteroduplex formation by recA protein: polarity of strand exchanges. Proc Natl Acad Sci USA 78:6149–6153

    Google Scholar 

  • Wolffe AP, Drew HR (1989) Initiation of transcription on nucleosomal templates. Proc Natl Acad Sci USA 86:9817–9821

    Google Scholar 

  • Wong EA, Capecchi MR (1987) Homologous recombination between coinjected DNA sequences peaks in early to mid-S phase. Mol Cell Biol 7:2294–2295

    Google Scholar 

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Communicated by M. Sekiguchi

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Kotani, H., Kmiec, E.B. DNA cruciforms facilitate in vitro strand transfer on nucleosomal templates. Molec. Gen. Genet. 243, 681–690 (1994). https://doi.org/10.1007/BF00279578

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  • DOI: https://doi.org/10.1007/BF00279578

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