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The replication behavior of Saccharomyces cerevisiae DNA in human cells

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Abstract

We studied the replication of random genomic DNA fragments from Saccharomyces cerevisiae in a long-term assay in human cells. Plasmids carrying large yeast DNA fragments were able to replicate autonomously in human cells. Efficiency of replication of yeast DNA fragments was comparable to that of similarly sized human DNA fragments and better than that of bacterial DNA. This result suggests that yeast genomic DNA contains sequence information needed for replication in human cells. To examine whether DNA replication in human cells would initiate specifically at a yeast origin of replication, we monitored initiation on a plasmid containing the yeast 2-micron autonomously replicating sequence (ARS) in yeast and human cells. We found that while replication initiates at the 2-micron ARS in yeast, it does not preferentially initiate at the ARS in human cells. This result suggests that the sequences that direct site specific replication initiation in yeast do not function in the same way in human cells, which initiate replication at a broader range of sequences.

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References

  • Bell SP, Stillman B (1992) ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex. Nature 357:128–134

    Google Scholar 

  • Brewer BJ, Fangman WL (1987) The localization of replication origins on ARS plasmids in S. cerevisiae. Cell 51:463–471

    Google Scholar 

  • Burhans WC, Vassilev LT, Caddle MS, Heintz NH, DePamphilis ML (1990) Identification of an origin of bidirectional DNA replication in mammalian chromosomes. Cell 62:955–965

    Google Scholar 

  • Diffley JFX, Cocker JH (1992) Protein-DNA interactions at a yeast replication origin. Nature 357:169–172

    Google Scholar 

  • Gahn TA, Schildkraut CL (1989) The Epstein-Barr virus origin of plasmid replication, PitoriP, contains both the initiation and termination sites of DNA replication. Cell 58:527–535

    Google Scholar 

  • Haase SB, Calos MP (1991) Replication control of autonomously replicating sequences. Nucleic Acids Res 19:5035–5058

    Google Scholar 

  • Handeli S, Klar A, Meuth M, Cedar H (1989) Mapping replication units in animal cells. Cell 57:909–920

    Google Scholar 

  • Harland RM, Laskey RA (1980) Regulated replication of DNA microinjected into eggs of X. laevis. Cell 21:761–771

    Google Scholar 

  • Heck MMS, Spradling AC (1990) Multiple replication origins are used during Drosophila chorion gene amplification. J Cell Biol 110:903–914

    Google Scholar 

  • Heintz NH, Hamlin JL (1982) An amplified chromosomal sequence that includes the gene for dihydrofolate reductase initiates replication within specific restriction fragments. Proc Natl Acad Sci USA 79:4083–4087

    Google Scholar 

  • Heizel SS, Krysan PJ, Tran CT, Calos MP (1991) Autonomous DNA replication in human cells is affected by the size and the source of the DNA. Mol Cell Biol 11:2263–2272

    Google Scholar 

  • Hirt B (1967) Selective extraction of polyoma DNA from infected mouse cultures. J Mol Biol 26:365–369

    Google Scholar 

  • Huberman JA (1991) Cell cycle control of initiation of eukaryotic DNA replication. Chromosoma 100:419–423

    Google Scholar 

  • Huberman JA, Spotila LD, Nawotka KA, El-Assouli SM, Davis LR (1987) The in vivo replication origin of yeast 2 μm plasmid. Cell 51:473–481

    Google Scholar 

  • Huberman JA, Zhu J, Davis LR, Newlon CS (1988) Close association of a DNA replication origin and an ARS element on chromosome III of the yeast, Saccharomyces cerevisiae. Nucleic Acids Res 16:6373–6384

    Google Scholar 

  • Hyrien O, Mechali M (1992) Plasmid replication in Xenopus eggs and egg extracts: 2D gel electrophoretic analysis. Nucleic Acids Res 20:1463–1469

    Google Scholar 

  • Krysan P, Calos MP (1991) Replication initiates at multiple locations on an autonomously replicating plasmid in human cells. Mol Cell Biol 11:1464–1472

    Google Scholar 

  • Krysan PJ, Haase SB, Calos MP (1989) Isolation of human sequences that replicate autonomously in human cells. Mol Cell Biol 9:1026–1033

    Google Scholar 

  • Linskens MHK, Huberman JA (1988) Organization of replication of ribosomal DNA in Saccharomyces cerevisiae. Mol Cell Biol 8:4927–4935

    Google Scholar 

  • Mahbubani HM, Paull T, Elder JK, Blow JJ (1992) DNA replication initiates at multiple sites on plasmid DNA in Xenopus egg extracts. Nuclei Acids Res 20:1457–1462

    Google Scholar 

  • Mechali M, Kearsey S (1984) Lack of specific sequence requirement for DNA replication in Xenopus eggs comared with high sequence specificity in yeast. Cell 38:55–64

    Google Scholar 

  • Newlon CS (1988) Yeast chromosome replication and segregation. Microbiol Rev 54:568–601

    Google Scholar 

  • Shinomiya T, Ina S (1991) Analysis of chromosomal replicons in early embryos of Drosophila melanogaster by two-dimensional gel electrophoresis. Nucleic Acdis Res 19:3935–3941

    Google Scholar 

  • Stillman BW, Gluzman Y (1985) Replication and supercoiling of simian virus 40 DNA in cell extracts from human cells. Mol Cell Biol 5:2051–2060

    Google Scholar 

  • Vassilev LT, Burhans WC, DePamphilis ML (1990) Mapping an origin of DNA replication at a single-copy locus in exponentially proliferating mammalian cell. Mol Cell Biol 10:4685–4689

    Google Scholar 

  • Vaughn JP, Dijkwel PA, Hamlin JL (1990) Replication initiates in a broad zone in the amplified CHO dihydrofolate reductase domain. Cell 61:1075–1087

    Google Scholar 

  • Wigler M, Sweet R, Sim GK, Wold B, Pellicer A, Lacy E, Maniatis T, Silverstein S, Axel R (1979) Transformation of mammalian cells with genes from prokaryotes and eukaryotes. Cell 16:777–785

    Google Scholar 

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by J.A. Huberman

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Tran, C.T., Caddle, M.S. & Calos, M.P. The replication behavior of Saccharomyces cerevisiae DNA in human cells. Chromosoma 102, 129–136 (1993). https://doi.org/10.1007/BF00356030

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