Abstract
Topoisomerase I is believed to be sufficient for early replication of circular viral genomes such as those of SV40 and of yeast plasmids. Topoisomerase II is required for the decatenation of the daughter genomes and probably also for fork elongation during the later stages of SV40 replication. Using the neutral-neutral two-dimensional gel system, we have followed the progression of replication of both SV40 and the yeast 2μm plasmid under various conditions of topoisomerase inhibition. During SV40 replication, inhibition of topoisomerase II by VP16, VM26 or hypertonic shock (but not by merbarone), and inhibition of topoisomerase I by camptothecin all led to the accumulation of aberrant DNA structures containing two almost completely replicated genomes. These aberrant structures resembled either recombination intermediates or late Cairns structures in which the site of replication termination had shifted and now mapped to a continuum of sites throughout the genome. Replication of the 2 μm plasmid in a topoisomerase II- but not a topoisomerase I-deficient yeast gave rise to very similar structures. The data suggest that inactivation of topoisomerase I or II either stimulates recombination or, by differentially affecting replication fork progression, leads to aberrant replication termination.
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References
Avemann K, Knippers R, Koller T, Sogo JM (1988) Camptothecin, a specific inhibitor of type I DNA topoisomerase, induces DNA breakage at replication forks. Mol Cell Biol 8:3026–3034
Bell L, Byers B (1979) Occurence of crossed strand-exchange forms in yeast DNA during meiosis. Proc Natl Acad Sci (USA) 76:3445–3449
Bell L, Byers B (1983) Separation of branched from linear DNA by two-dimensional gel electrophoresis. Anal Biochem 130:527–535
Brewer BJ, Fangman WL (1987) The localization of replication origins on ARS plasmids inS. cerevisiae. Cell 51:463–471
BrewerBJ, Fangman WL (1988) A replication fork barrier at the 3′ end of yeast ribosomal RNA genes. Cell 55:637–643
Brewer BJ, Fangman WL (1993) Initiation at closely spaced replication origins in a yeast chromosome. Science 262:1728–1731
Brewer BJ, Fangman WL (1994) Initiation preference at a yeast origin of replication. Proc Natl Acad Sci USA 91:3418–3422
Brewer BJ, Lockshon D, Fangman WL (1992) The arrest of replication forks in the rDNA of yeast occurs independently of transcription. Cell 71:267–276
Brill SJ, DiNardo S, Voelkel-Meiman K, Sternglanz R (1987) Need for DNA topoisomerase activity as a swivel for DNA replication for transcription of ribosomal RNA. Nature 326:414–416
Brockman WW, Gutai MW, Nathans D (1975) Evolutionary variants of simian virus 40: characterisation of cloned complementing variants. Virology 66:36–52
Caddle MS, Calos MP (1994) Specific initiation at an origin of replication fromSchizosaccharomyces pombe. Mol Cell Biol 14:1796–1805
Charron M, Hancock R (1990) Roles of DNA topoisomerases I and II in DNA replication, mitotic chromosome formation, and recombination in mammalian cells. In: Harris JR, Zbarsky IB (eds) Nuclear structure and function. Plenum Press, New York, pp 405–411
Charron M, Hancock R (1991) Chromosome recombination and defective genome segragation induced in chinese hamster cells by the topoisomerase II inhibitor VM-26. Chromosoma 100:97–102
Delidakis C, Kafatos FC (1989) Amplification enhancers and replication origins in the autosomal chorion gene cluster ofDrosophila. EMBO J 8:891–901
Depamphilis ML, Bradley MK (1986) Replication of simian virus 40 and polyoma virus genomes. In: Salzman NP (ed) The Papovaviridae, vol 1, Plenum Press, New York, pp 99–246
Dijkwel PA, Vaughn JP, Hamlin JL (1991) Mapping of replication initiation sites in mammalian genomes by two-dimensional gel analysis: stabilization and enrichment of replication intermediates by isolation on the nuclear matrix. Mol Cell Biol 11:3850–3859
DiNardo S, Voelkel K, Sternglanz R (1984) DNA topoisomerase II mutant ofSaccharomyces cerevisiae: topoisomerase II is required for segregation of daughter molecules at the termination of DNA replication. Proc Natl Acad Sci USA 81:2616–2620
Drake FH, Hofmann GA, Mong S-M, Bartus JO, Hertzberg RP, Johnson RK, Mattern MR, Mirabelli CK (1989) In vitro and intracellular inhibition of topoisomerase II by the antitumor agent Merbarone. Cancer Res 49:2578–2583
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Feinberg AP, Vogelstein B (1984) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Addendum. Anal Biochem 137:266–267
Gahn TA, Schildkraut CL (1989) The Epstein-Barr virus origin of plasmid replication, oriP, contains both the initiation and termination sites of DNA replication. Cell 58:527–535
Heck MMS, Spradling AC (1990) Multiple replication origins are used duringDrosophila chorion gene amplification. J Cell Biol 110:903–914
Hernández P, Mart’n-Parras L, Mart’nez-Robles ML, Schvartzman JB (1993) Conserved features in the mode of replication of eukaryotic ribosomal RNA genes. EMBO J 12:1475–1485
Hirt B (1967) Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol 26:365–369
Holm C, Goto T, Wang JC, Botstein D (1985) DNA topoisomerase II is required at the time of mitosis in yeast. Cell 41:553–563
Huberman JA, Spotila LA, Nawotka KA, El-Assouli SM, Davis LR (1987) The in vivo replication origin of the yeast 2 μm plasmid. Cell 51:473–481
Ishimi Y, Sugasawa K, Hanaoka F, Eki T, Hurwitz J (1992) Topoisomerase II plays an essential role as a swivelase in the late stage of SV40 chromosome replication in vitro. J Biol Chem 267:462–466
Kim RA, Wang JC (1989) Function of DNA topoisomerases as replication swivels inSaccharomyces cerevisiae. J Mol Biol 208:257–267
Krysan PJ, Calos MP (1991) Replication initiates at multiple locations on an autonomously replicating plasmid in human cells. Mol Cell Biol 11:1464–1472
Lai C-J, Nathans D (1975) Non-specific termination of simian virus 40 DNA replication. J Mol Biol 97:113–118
Lee MP, Hsieh T (1992) Incomplete reversion of double stranded DNA cleavage mediated byDrosophila topoisomerase II: formation of single stranded DNA cleavage complex in the presence of an anti-tumor drug VM26. Nucleic Acids Res 20:5027–5033
Lim M, Liu LF, Jacobson-Kram D, Williams D (1986) Induction of sister chromatid exchanges by inhibitors of topoisomerases. Cell Biol Toxicol 2:485–496
Linskens MHK, Huberman JA (1988) Organization of replication of ribosomal DNA inSaccharomyces cerevisiae. Mol Cell Biol 8:4927–4935
Linskens MHK, Huberman JA (1990) Ambiguities in results obtained with 2D gel replicon mapping techniques. Nucleic Acids Res 18:647–652
Liu LF (1989) DNA topoisomerase poisons as antitumor drugs. Annu Rev Biochem 58:351–375
Mart’n-Parras L, Hernández P, Mart’nez-Robles ML, SchvartzmanJB (1991) Unidirectional replication as visualized by two-dimensional agarose gel electrophoresis. J Mol Biol 220:843–853
Oppenheim A (1981) Separation of closed circular DNA from linear DNA by electrophoresis in two dimensions in agarose gels. Nucleic Acids Res 9:6805–6812
Reynolds AE, McCarroll RM, Newlon CS, Fangman WL (1989) Time of replication of ARS elements along yeast chromosome III. Mol Cell Biol 9:4488–4494
Richter A, Strausfeld U (1988) Effects of VM26, a specific inhibitor of type II DNA topoisomerase, on SV40 chromatin replication in vitro. Nucleic Acids Res 21:10119–10129
Richter A, Strausfeld U, Knippers R (1987) Effects of VM26 (teniposide), a specific inhibitor of type II DNA topoisomerase, on SV40 DNA replication in vivo. Nucleic Acids Res 15:3455–3468
Shinomiya T, Ina S (1993) DNA replication of histone gene repeats inDrosophila melanogaster tissue culture cells: multiple initiation sites and replication pause sites. Mol Cell Biol 13:4098–4106
Snapka RM (1986) Topoisomerase inhibitors can selectively interfere with different stages of simian virus 40 DNA replication. Mol Cell Biol 6:4221–4227
Snapka RM, Permana PA (1993) Sv40 DNA replication intermediates: analysis of drugs which target mammalian DNA replication. Bioessays 15:121–127
Snapka RM, Powelson MA, Strayer JM (1988) Swiveling and decatenation of replicating simian virus 40 genomes in vivo. Mol cell Biol 8:515–521
Sundin O, Varshavsky A (1980) Terminal stages of SV40 DNA replication proceed via multiply intertwined catenated dimers. Cell 21:103–114
Sundin O, Varshavsky A (1981) Arrest of segregation leads to accumulation of highly intertwined catenated dimers: dissection of the final stages of SV40 DNA replication. Cell 25:659–669
Tapper DP, Depamphilis ML (1978) Discontinuous DNA replication: accumulation of simian virus 40 DNA at specific stages in its replication. J Mol Biol 120:401–422
Thrash C, Voelkel K, DiNardo S, Sternglanz R (1984) Identification ofSaccharomyces cerevisiae mutants deficient in DNA topoisomerase I activity. J Biol Chem 259:1375–1377
Tsao YP, Russo A, Nyamuswa G, Silber R, Liu LF (1993) Interaction between replication forks and topoisomerase I-DNA cleavable complexes: studies in a cell-free SV40 DNA replication system. Cancer Res 53:5908–5914
Uemura T, Ohkura H, Adachi Y, Morino K, Shiozaki K, Yanagida M (1987) DNA topoisomerase II is required for condensation and separation of mitotic chromosomes inS. pombe. Cell 50:917–925
Umek RM, Linskens MHK, Kowalski D, Huberman JA (1989) New beginnings in studies of eukaryotic DNA replication origins. Biochim Biophys Acta 1007:1–14
Vaughn JP, Dijkwel PA, Hamlin JL (1990) Replication initiates in a broad zone in the amplified CHO dihydrofolate reductase domain. Cell 61:1075–1087
Wang JC (1985) DNA topoisomerases. Annu Rev Biochem 54:665–697
Weaver DT, Fields-Berry SC, Depamphilis ML (1985) The termination region for SV40 DNA replication directs the mode of separation for the two sibling molecules. Cell 41:565–575
Yang L, Wold MS, Li JJ, Kelly TJ, Liu LF (1987) Roles of DNA topoisomerases in simian virus 40 DNA replication in vitro. Proc Natl Acad Sci USA 84:950–954
Zhu J, Newlon CS, Huberman JA (1992) Localization of a DNA replication origin and termination zone on chromosome III ofSaccharomyces cerevisiae. Mol Cell Biol 12:4733–4741
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Edited by: J. Huberman
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Levac, P., Moss, T. Inactivation of topoisomerase I or II may lead to recombination or to aberrant replication termination on both SV40 and yeast 2 μm DNA. Chromosoma 105, 250–260 (1996). https://doi.org/10.1007/BF02528774
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DOI: https://doi.org/10.1007/BF02528774