Home LiteratureArticle Details
PMID: 6086310 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Localization of T-antigen on simian virus 40 minichromosomes by immunoelectron microscopy.

The EMBO journal ·Vol. 3 ·No. 6 ·1984-06-00 ·Pages 1235-41

Harper F, Florentin Y, Puvion E

Abstract

SV40 chromatin extracted from 42 h post-infected cells by a modification of the standard Triton X-100-EDTA procedure and purified on neutral sucrose gradients was partially immunoprecipitable by a specific SV40 T-antigen (T-Ag) antiserum. Electron microscopic observations of spread minichromosomes were made after labelling by the indirect colloidal gold immunological method using monoclonal antibodies specific for the SV40 T-Ag. In 1-2% of morphologically mature minichromosomes the labelling corresponding to tightly bound T-Ag was localized within the nucleosome-free region near one of its borders. Mapping with three single-cut restriction endonucleases: BamHI, EcoRI and BglI localized the labelling near to, or at the origin of, replication. In addition, we observed that the T-Ag specific antibodies were linked to a DNA-bound particle when the region was not masked by a large clump of antibodies. The variable size of this particle led us to suggest that it might be a complex of T-Ag with other proteins.

MeSH Terms
Animals Antibodies, Monoclonal Antigen-Antibody Complex/analysis Antigens, Polyomavirus Transforming Antigens, Viral, Tumor/genetics Cell Line Chlorocebus aethiops Chromatin/ultrastructure Chromosomes/ultrastructure Immune Sera Kidney Microscopy, Electron Protein Kinases/genetics Simian virus 40/enzymology,genetics,ultrastructure Viral Proteins/genetics
Chemicals
Antibodies, Monoclonal Antigen-Antibody Complex Antigens, Polyomavirus Transforming Antigens, Viral, Tumor Chromatin Immune Sera Viral Proteins Protein Kinases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Harper F
Florentin Y
Puvion E
References (52)
52 references, click to expand
  1. Mutational analysis of the simian virus 40 replicon: pseudorevertants of mutants with a defective replication origin.
    Proc Natl Acad Sci U S A. 1979 Dec;76(12):6128-31 PMID: 230499
  2. Interaction of Simian Virus 40 chromatin with Simian Virus 40 T-antigen.
    J Virol. 1977 Nov;24(2):451-60 PMID: 199733
  3. Analysis of simian virus 40 chromosome-T-antigen complexes: T-antigen is preferentially associated with early replicating DNA intermediates.
    J Virol. 1983 Oct;48(1):281-95 PMID: 6310148
  4. The SV40 early region TATA box is required for accurate in vitro initiation of transcription.
    Nature. 1981 Mar 26;290(5804):310-5 PMID: 6259539
  5. Compact form of SV40 viral minichromosome is resistant to nuclease: possible implications for chromatin structure.
    Nucleic Acids Res. 1977 Oct;4(10):3303-25 PMID: 200892
  6. Mapping the in vivo arrangement of nucleosomes on simian virus 40 chromatin by the photoaddition of radioactive hydroxymethyltrimethylpsoralen.
    J Virol. 1982 Jan;41(1):78-87 PMID: 6283130
  7. SV40 viral minichromosome: preferential exposure of the origin of replication as probed by restriction endonucleases.
    Nucleic Acids Res. 1978 Oct;5(10):3469-77 PMID: 214758
  8. Absence of nucleosomes in a fraction of SV40 chromatin between the origin of replication and the region coding for the late leader RNA.
    Cell. 1980 May;20(1):65-73 PMID: 6248237
  9. Subunit structure of simian-virus-40 minichromosome.
    Eur J Biochem. 1976 Nov 15;70(2):543-53 PMID: 188649
  10. Autoregulation of simian virus 40 gene A by T antigen.
    Proc Natl Acad Sci U S A. 1976 Sep;73(9):3083-7 PMID: 184459
  11. N-Acetoxy-acetylaminofluorene reacts preferentially with a control region of intracellular SV40 chromosome.
    Nature. 1981 May 7;291(5810):84-5 PMID: 6262649
  12. Association of simian virus 40 T antigen with simian virus 40 nucleoprotein complexes.
    J Virol. 1979 Jan;29(1):232-41 PMID: 219219
  13. Chromatin structure: deduced from a minichromosome.
    Science. 1975 Mar 28;187(4182):1202-3 PMID: 17754289
  14. Simian virus 40-transformed cells express new species of proteins precipitable by anti-simian virus 40 tumor serum.
    J Virol. 1979 Aug;31(2):472-83 PMID: 225566
  15. Chromatin-like structures in polyoma virus and simian virus 10 lytic cycle.
    J Virol. 1975 Jan;17(1):204-11 PMID: 173884
  16. Visualization of anti-histone antibodies on SV40 minichromosomes by scanning transmission electron microscopy (STEM).
    Exp Cell Res. 1983 Aug;147(1):135-42 PMID: 6311595
  17. A high-resolution electron microscopy study of nucleosomes from simian virus 40 chromatin.
    J Mol Biol. 1981 Jul 15;149(4):735-44 PMID: 6273589
  18. A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels.
    Anal Biochem. 1979 Sep 15;98(1):231-7 PMID: 94518
  19. Electron microscope localization of a protein bound near the origin of simian virus 40 DNA replication.
    J Virol. 1975 Jan;15(1):167-72 PMID: 163341
  20. T antigen repression of SV40 early transcription from two promoters.
    Cell. 1981 Dec;27(3 Pt 2):603-13 PMID: 6101224
  21. SV40 T antigen binding site mutations that affect autoregulation.
    Cell. 1983 Apr;32(4):1227-40 PMID: 6301686
  22. The binding site on SV40 DNA for a T antigen-related protein.
    Cell. 1978 Jan;13(1):165-79 PMID: 202398
  23. Mapping of SV40 DNA replication origin region binding sites for the SV40 T antigen by protection against exonuclease III digestion.
    Cell. 1980 Jun;20(2):411-22 PMID: 6248243
  24. Origin of DNA replication in papovavirus chromatin is recognized by endogenous endonuclease.
    Proc Natl Acad Sci U S A. 1978 Dec;75(12):5964-8 PMID: 216004
  25. Higher order structure of simian virus 40 chromatin.
    Science. 1978 Aug 4;201(4354):406-15 PMID: 208155
  26. Heterogeneity in the interaction of chromatin subunits with anti-histone sera visulatized by immuno-electron microscopy.
    Exp Cell Res. 1978 Mar 1;112(1):1-14 PMID: 75806
  27. Specific association of simian virus 40 tumor antigen with simian virus 40 chromatin.
    J Virol. 1980 Jan;33(1):78-87 PMID: 6245256
  28. Alternative interactions of the SV40 A protein with DNA.
    Virology. 1981 Nov;115(1):75-87 PMID: 6270902
  29. Protein-SV40 DNA complex stable in high salt and sodium dodecyl sulfate.
    Biochem Biophys Res Commun. 1976 Feb 9;68(3):927-36 PMID: 177014
  30. Simian virus 40 early mRNA's contain multiple 5' termini upstream and downstream from a Hogness-Goldberg sequence; a shift in 5' termini during the lytic cycle is mediated by large T antigen.
    J Virol. 1981 Oct;40(1):224-40 PMID: 6270376
  31. Characterization of the autoregulation of simian virus 40 gene A.
    J Virol. 1977 Oct;24(1):22-7 PMID: 198575
  32. Protein-DNA interactions at the origin of simian virus 40 DNA replication.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 2:655-61 PMID: 226310
  33. Identification of regions of the SV40 genome which contain preferred SV40 T antigen-binding sites.
    Cell. 1976 Aug;8(4):535-45 PMID: 182383
  34. In vivo sequence requirements of the SV40 early promotor region.
    Nature. 1981 Mar 26;290(5804):304-10 PMID: 6259538
  35. Relationship of replication and transcription of Simian Virus 40 DNA.
    Proc Natl Acad Sci U S A. 1973 Jul;70(7):1927-30 PMID: 4352963
  36. Simian virus 40 large tumor antigen on replicating viral chromatin: tight binding and localization on the viral genome.
    J Virol. 1983 Jul;47(1):65-76 PMID: 6306282
  37. Viral DNA synthesis in cells infected by temperature-sensitive mutants of simian virus 40.
    J Virol. 1974 Jul;14(1):116-24 PMID: 4366021
  38. Simian virus 40 deoxyribonucleic acid synthesis: the viral replicon.
    J Virol. 1972 Oct;10(4):591-8 PMID: 4343541
  39. Sites in simian virus 40 chromatin which are preferentially cleaved by endonucleases.
    Cell. 1978 Dec;15(4):1511-8 PMID: 215331
  40. Intracellular SV40 nucleoprotein complexes: synthesis to encapsidation.
    Virology. 1980 Dec;107(2):389-401 PMID: 6256939
  41. SV40 gene expression is modulated by the cooperative binding of T antigen to DNA.
    Cell. 1981 Aug;25(2):373-84 PMID: 6269743
  42. A new preparation method for dark-field electron microscopy of biomacromolecules.
    J Ultrastruct Res. 1971 Apr;35(1):147-67 PMID: 4931423
  43. Salt-resistant association of simian virus 40 T antigen with simian virus 40 DNA in nucleoprotein complexes.
    J Virol. 1983 Aug;47(2):276-86 PMID: 6312066
  44. Binding of an SV40 T antigen-related protein to the DNA of SV40 regulatory mutants.
    Nature. 1981 Feb 26;289(5800):810-3 PMID: 6258090
  45. Oligomeric structure of a simian virus 40 T antigen in free form and bound to DNA.
    J Mol Biol. 1981 Jun 5;148(4):347-53 PMID: 6273580
  46. Monoclonal antibodies specific for simian virus 40 tumor antigens.
    J Virol. 1981 Sep;39(3):861-9 PMID: 6169844
  47. Folding of the DNA double helix in chromatin-like structures from simian virus 40.
    Proc Natl Acad Sci U S A. 1975 May;72(5):1843-7 PMID: 168578
  48. Association of simian virus 40 T antigen with replicating nucleoprotein complexes of simian virus 40.
    J Virol. 1980 Aug;35(2):320-30 PMID: 6255173
  49. A nucleosome-free region in SV40 minichromosomes.
    Nature. 1980 May 22;285(5762):263-5 PMID: 6246449
  50. Nucleosome structure studied with purified antibodies to histones H2B, H3 and H4.
    FEBS Lett. 1977 Dec 20;85(1):61-4 PMID: 598520
  51. A stretch of "late" SV40 viral DNA about 400 bp long which includes the origin of replication is specifically exposed in SV40 minichromosomes.
    Cell. 1979 Feb;16(2):453-66 PMID: 222461
  52. Topography of simian virus 40 A protein-DNA complexes: arrangement of protein bound to the origin of replication.
    J Virol. 1983 Apr;46(1):151-61 PMID: 6298452
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1984-06-00
Pages
1235-41
Language
English
Region
England
NLM ID
8208664
PMCID
PMC557504
Subset
IM
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2