Skip to main content
SpringerLink
Log in

Replication of SV40 minichromosomes in vitro

  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

We operationally define two forms of SV40 minichromosomes, a 75S-form, prepared at low salt concentration, referred to as native minichromosomes, and a 50S-form, obtained after treatment with 0.5M potassium acetate, the salt-treated minichromosomes. Both preparations of minichromosomes serve well as templates for replication in vitro. Their respective replication products are strikingly different: replicated native minichromosomes contain a densely packed array of the maximal number of nucleosomes whereas replicated salt-treated minichromosomes carry, on average, half of the maximal number. We conclude that in both cases parental nucleosomes are transferred to progeny DNA, and, in addition, that an assembly of new nucleosomes occurs during the replication of native minichromosomes. This is apparently due to the presence of a nucleosome assembly factor as a constituent of native minichromosomes that dissociates upon treatment with salt. We further show that preparations of minichromosomes usually contain significant amounts of copurifying hnRNP particles and SV40 virion precursor particles. However, these structures do not detectably affect the replication and the chromatin assembly reactions.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Baumgartner I, Kuhn C, Fanning E (1979) Identification and characterization of fast-sedimenting SV40 nucleoprotein complexes. Virology 96: 54–63

    Article  PubMed  CAS  Google Scholar 

  • Beyer AL, Christensen ME, Walker BW, LeStourgeon WM (1977) Identification and characterization of the packaging proteins of core 40S hnRNP particles. Cell 11: 127–138

    Article  PubMed  CAS  Google Scholar 

  • Bonne-Andrea C, Wong ML, Alberts B (1990) In vitro replication through nucleosomes without histone displacement. Nature(London) 343: 719–726

    CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein dye binding. Anal Biochem 72: 248–254

    Article  PubMed  CAS  Google Scholar 

  • Challberg MD, Kelly TJ (1989) Animal virus DNA replication. Ann Rev Biochem 58: 671–717

    Article  PubMed  CAS  Google Scholar 

  • Cusick ME, DePamphilis ML, Wassarman (1984) Dispersive segregation of nucleosomes during replication of SV40 chromosomes. J Mol Biol 178: 249–271

    Article  PubMed  CAS  Google Scholar 

  • Dreyfuss G (1986) Structure and function of nuclear and cytoplasmic ribonucleoprotein particles. Ann Rev Cell Biol 2: 459–498

    PubMed  CAS  Google Scholar 

  • Dreyfuss G, Swanson MS, Pinol-Roma S (1988) Heterogeneous nuclear ribonucleoprotein particles and the pathway of mRNA formation. Trends in Biochem Sci 13: 86–90

    Article  CAS  Google Scholar 

  • DePamphilis ML, Bradley MK (1986) Replication of SV40 and polyoma chromosomes. In: Saltzman NP (ed) The Papovaviridae. Plenum Publishing Corp., New York, pp 99–246

    Google Scholar 

  • Germond, JE, Hirt B, Oudet P, Gross-Bellard M, Chambon P (1975) Folding of the DNA double helix in a chromatin-like structure from Simian Virus 40. Proc Natl Acad Sci USA 72: 1843–1847

    Article  PubMed  CAS  Google Scholar 

  • Herman T, DePamphilis ML, Wassarman PM (1981) Structure of chromatin at desoxyribonucleic acid replication forks: location of the first nucleosomes on newly synthesized simian virus 40 desoxyribonucleic acid. biochemistry 20: 621–630

    Article  PubMed  CAS  Google Scholar 

  • Ishimi Y, Sugasawa K, Hanaoka F, Kikuchi A (1991) Replication of the Simian Virus 40 chromosome with purified proteins. J Biol Chem 266: 16141–16148

    PubMed  CAS  Google Scholar 

  • Jackson V (1990) In vivo studies on the dynamics of histone-DNA interaction: Evidence for nucleosome dissolution during replication and transcription and a low level of dissolution independent of both. Biochemistry 29: 719–731

    Article  PubMed  CAS  Google Scholar 

  • Jakobovitz EG, Bratosin G, Aloni Y (1980) A nucleosome-free region in SV40 minichromosomes Nature (London) 285: 263–265

    Article  Google Scholar 

  • Klempnauer KH, Fanning E, Otto B, Knippers R (1980) Maturation of newly replicated chromatin of simian virus 40 and its host cell. J Mol Biol. 136: 359–374

    Article  PubMed  CAS  Google Scholar 

  • Krude T, Knippers R (1991) Transfer of nucleosomes from parental to replicated chromatin. Mol Cell Biol 11: 6257–6267.

    PubMed  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227: 680–685

    Article  CAS  Google Scholar 

  • Li JJ, Kelly TJ (1984) Simian virus 40 DNA replication in vitro. Proc Natl Acad Sci USA 81: 6973–6977.

    Article  PubMed  CAS  Google Scholar 

  • Müller U, Zentgraf H, Eicken I, Keller W (1978) Higher order structure of simian virus 40 chromatin. Science 201: 406–415

    PubMed  Google Scholar 

  • Otto B, Böhm J, Knippers R (1980) A histone-specific acetyltransferase is associated with simian-virus-40 chromatin. Eur J Biochem 112: 363–366

    Article  PubMed  CAS  Google Scholar 

  • Pülm W, Knippers R (1984) Chromatin structure and DNA replication. In: Proteins involved in DNA replication. Hübscher U, Spadari S (eds) Plenum Press, New York and London

    Google Scholar 

  • Randall SK, Kelly TJ (1992) The fate of parental nucleosomes during SV40 DNA replication. J Biol Chem 267: 14259–14265

    PubMed  CAS  Google Scholar 

  • Saragosti S, Moyne G, Yaniv M (1980) Absence of nucleosomes in a fraction of SV40 chromatin between the origin of replication and the region coding for the late leader RNA. Cell 20: 65–73

    Article  PubMed  CAS  Google Scholar 

  • Schlaeger EJ, Knippers R (1979) DNA-histone interactions in the vicinity of replication points. Nucleic Acids Res 6: 645–656

    PubMed  CAS  Google Scholar 

  • Schlaeger EJ, Pülm W, Knippers R (1983) Chromatin maturation depends on continued DNA replication. FEBS Lett 156: 281–286

    Article  PubMed  CAS  Google Scholar 

  • Sheinin R, Humbert J, Pearlman RE (1978) Some aspects of eukaryotic DNA replication. Ann Rev Biochem 47: 277–316

    Article  PubMed  CAS  Google Scholar 

  • Smith S, Stillman BW (1989) Purification and characterization of CAF-I, a human cell factor required for chromatin assembly during DNA replication in vitro. Cell 58: 15–25

    Article  PubMed  CAS  Google Scholar 

  • Smith S, Stillman BW (1991a) Stepwise assembly of chromatin during DNA replication in vitro. EMBO J 10: 971–980

    PubMed  CAS  Google Scholar 

  • Smith S, Stillman BW (1991b) Immunological characterization of chromatin assembly factor I, a human cell factor required for chromatin assembly during DNA replication in vitro. J Biol Chem 266: 12041–12047

    PubMed  CAS  Google Scholar 

  • Sogo JM, Stahl H, Koller T, Knippers R (1986) Structure of replicating simian virus 40 minichromosomes. J Mol Biol 189: 189–204

    Article  PubMed  CAS  Google Scholar 

  • Stillman BW (1989) Initiation of eukaryotic DNA replication in vitro. Ann Rev Cell Biol 5: 197–245

    PubMed  CAS  Google Scholar 

  • Su R, DePamphilis ML (1978). Simian virus 40 DNA replication in isolated replicating viral chromosomes. J Virol 28: 53–65

    PubMed  CAS  Google Scholar 

  • Sugasawa K, Murakami Y, Miyamoto N, Hanaoka F, Ui M (1990) Assembly of nascent DNA into nucleosome structures in simian virus 40 chromosomes by HeLa cell extract. J Virol 64: 4820–4829

    PubMed  CAS  Google Scholar 

  • Van Holde KE (1989). Chromatin. Springer, New York Heidelberg Berlin

    Google Scholar 

  • Wray W, Boulikas T, Wray VP, Hancock R (1981) Silver staining of proteins in polyacrylamide gels. Anal Biochem 118: 197–203

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Fakultät für Biologie, Universität Konstanz, W-7750, Konstanz, Federal Republic of Germany

    Torsten Krude & Rolf Knippers

Authors
  1. Torsten Krude
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rolf Knippers
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Krude, T., Knippers, R. Replication of SV40 minichromosomes in vitro. Chromosoma 102 (Suppl 1), S83–S92 (1992). https://doi.org/10.1007/BF02451790

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02451790

Keywords

Search

Navigation