Home LiteratureArticle Details
PMID: 9811771 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The origin DNA-binding and single-stranded DNA-binding domains of simian virus 40 large T antigen are distinct.

Journal of virology ·Vol. 72 ·No. 12 ·1998-12-00 ·Pages 10256-9

Wu C, Edgil D, Simmons DT

Abstract

Little is known about the ability of simian virus 40 (SV40) T antigen to bind single-stranded DNA. We demonstrate here that a mutant (259-708) missing the first 258 amino acids of T antigen and its origin-binding domain bound single-stranded DNA at close to normal levels, whereas a mutant containing only the first 259 amino acids failed to bind any single-stranded DNA. The 259-708 mutant also assembled into high-molecular-weight oligomers in the presence of single-stranded DNA. Its ATPase activity was stimulated by single-stranded DNA similarly to the wild type (WT). Furthermore, WT T antigen's ability to bind to single-stranded DNA was inhibited by the binding of two monoclonal antibodies that recognize a region after residue 362. These results show that the domain responsible for binding to single-stranded DNA is completely separate from the origin-binding domain.

MeSH Terms
Adenosine Triphosphatases/metabolism Animals Antibodies, Monoclonal Antibodies, Viral Antigens, Polyomavirus Transforming/chemistry,genetics,metabolism Binding Sites/genetics DNA Replication DNA, Single-Stranded/genetics,metabolism DNA, Viral/genetics,metabolism Sequence Deletion
Chemicals
Antibodies, Monoclonal Antibodies, Viral Antigens, Polyomavirus Transforming DNA, Single-Stranded DNA, Viral Adenosine Triphosphatases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Wu C
Department of Biological Sciences, University of Delaware, Newark, Delaware 19716-2590, USA.
Edgil D
Simmons D T
References (50)
50 references, click to expand
  1. RNA unwinding activity of SV40 large T antigen.
    Cell. 1989 Jun 16;57(6):955-63 PMID: 2472217
  2. Simian virus 40 (SV40) large tumor antigen causes stepwise changes in SV40 origin structure during initiation of DNA replication.
    Proc Natl Acad Sci U S A. 1989 Jun;86(11):3939-43 PMID: 2542957
  3. Properties of the DNA-binding domain of the simian virus 40 large T antigen.
    Mol Cell Biol. 1989 Dec;9(12):5525-36 PMID: 2555700
  4. Four major sequence elements of simian virus 40 large T antigen coordinate its specific and nonspecific DNA binding.
    J Virol. 1990 May;64(5):1973-83 PMID: 2157865
  5. Mapping of helicase and helicase substrate-binding domains on simian virus 40 large T antigen.
    J Virol. 1990 May;64(5):2014-20 PMID: 2157869
  6. Identification of simian virus 40 T-antigen residues important for specific and nonspecific binding to DNA and for helicase activity.
    J Virol. 1990 Oct;64(10):4858-65 PMID: 2168972
  7. Interaction of DNA polymerase alpha-primase with cellular replication protein A and SV40 T antigen.
    EMBO J. 1992 Feb;11(2):769-76 PMID: 1311258
  8. Strand-specific recognition of a synthetic DNA replication fork by the SV40 large tumor antigen.
    Science. 1992 Jun 19;256(5064):1656-61 PMID: 1319087
  9. The simian virus 40 T antigen double hexamer assembles around the DNA at the replication origin.
    J Biol Chem. 1992 Jul 15;267(20):14129-37 PMID: 1321135
  10. Structure and function of simian virus 40 large tumor antigen.
    Annu Rev Biochem. 1992;61:55-85 PMID: 1323237
  11. Nonspecific DNA binding activity of simian virus 40 large T antigen: evidence for the cooperation of two regions for full activity.
    J Virol. 1992 Sep;66(9):5443-52 PMID: 1323705
  12. Initiation of simian virus 40 DNA replication requires the interaction of a specific domain of human DNA polymerase alpha with large T antigen.
    Mol Cell Biol. 1993 Feb;13(2):809-20 PMID: 8380896
  13. An interaction between replication protein A and SV40 T antigen appears essential for primosome assembly during SV40 DNA replication.
    J Biol Chem. 1993 Feb 15;268(5):3389-95 PMID: 8381428
  14. Recognition of model DNA replication forks by the SV40 large tumor antigen.
    Chromosoma. 1992;102(1 Suppl):S46-51 PMID: 1337879
  15. Biochemical analysis of mutants with changes in the origin-binding domain of simian virus 40 tumor antigen.
    J Virol. 1993 Jul;67(7):4227-36 PMID: 8389924
  16. Strand and face: the topography of interactions between the SV40 origin of replication and T-antigen during the initiation of replication.
    EMBO J. 1994 Feb 15;13(4):982-92 PMID: 8112311
  17. Replication protein A induces the unwinding of long double-stranded DNA regions.
    J Mol Biol. 1996 May 31;259(1):104-12 PMID: 8648638
  18. Purification of the simian virus 40 (SV40) T antigen DNA-binding domain and characterization of its interactions with the SV40 origin.
    J Virol. 1997 May;71(5):3972-85 PMID: 9094674
  19. Six molecules of SV40 large T antigen assemble in a propeller-shaped particle around a channel.
    J Mol Biol. 1997 Apr 25;268(1):15-20 PMID: 9149137
  20. Denaturation of the simian virus 40 origin of replication mediated by human replication protein A.
    Mol Cell Biol. 1997 Jul;17(7):3876-83 PMID: 9199322
  21. Dimerization of simian virus 40 T-antigen hexamers activates T-antigen DNA helicase activity.
    J Virol. 1997 Nov;71(11):8766-73 PMID: 9343236
  22. The initiation of simian virus 40 DNA replication in vitro.
    Crit Rev Biochem Mol Biol. 1997;32(6):503-68 PMID: 9444478
  23. Topoisomerase I stimulates SV40 T antigen-mediated DNA replication and inhibits T antigen's ability to unwind DNA at nonorigin sites.
    Virology. 1998 Mar 15;242(2):435-43 PMID: 9514975
  24. The activity of topoisomerase I is modulated by large T antigen during unwinding of the SV40 origin.
    J Biol Chem. 1998 Aug 7;273(32):20390-6 PMID: 9685392
  25. Studies on the initiation and elongation reactions in the simian virus 40 DNA replication system.
    Proc Natl Acad Sci U S A. 1990 Dec;87(24):9712-6 PMID: 2175912
  26. Differential induction of structural changes in the simian virus 40 origin of replication by T antigen.
    J Virol. 1991 Mar;65(3):1228-35 PMID: 1847451
  27. Simian virus 40 large T antigen untwists DNA at the origin of DNA replication.
    J Biol Chem. 1991 Mar 15;266(8):5062-71 PMID: 1848235
  28. Effects of T antigen and replication protein A on the initiation of DNA synthesis by DNA polymerase alpha-primase.
    Mol Cell Biol. 1991 Apr;11(4):2108-15 PMID: 1848671
  29. The zinc finger region of simian virus 40 large T antigen is needed for hexamer assembly and origin melting.
    J Virol. 1991 Jun;65(6):3167-74 PMID: 1851875
  30. Simian virus 40 T-antigen DNA helicase is a hexamer which forms a binary complex during bidirectional unwinding from the viral origin of DNA replication.
    J Virol. 1992 Feb;66(2):804-15 PMID: 1309914
  31. Simian virus 40 deoxyribonucleic acid synthesis: the viral replicon.
    J Virol. 1972 Oct;10(4):591-8 PMID: 4343541
  32. A poly(dT)-stimulated ATPase activity associated with simian virus 40 large T antigen.
    J Biol Chem. 1979 Sep 10;254(17):8113-6 PMID: 224046
  33. Use of monoclonal antibodies as probes of simian virus 40 T antigen ATPase activity.
    J Biol Chem. 1981 Nov 25;256(22):11854-8 PMID: 6170639
  34. Relationship of oligomerization to enzymatic and DNA-binding properties of the SV40 large T antigen.
    Cell. 1982 Jan;28(1):125-34 PMID: 6461415
  35. An immunoaffinity purification procedure for SV40 large T antigen.
    Virology. 1985 Jul 15;144(1):88-100 PMID: 2998049
  36. DNA helicase activity of SV40 large tumor antigen.
    EMBO J. 1986 Aug;5(8):1939-44 PMID: 3019672
  37. Simian virus 40 (SV40) DNA replication: SV40 large T antigen unwinds DNA containing the SV40 origin of replication.
    Proc Natl Acad Sci U S A. 1987 Jan;84(1):16-20 PMID: 3025851
  38. Initiation of simian virus 40 DNA replication in vitro: large-tumor-antigen- and origin-dependent unwinding of the template.
    Proc Natl Acad Sci U S A. 1987 Jun;84(11):3643-7 PMID: 3035543
  39. Studies on the origin-specific DNA-binding domain of simian virus 40 large T antigen.
    J Virol. 1987 Oct;61(10):3326-30 PMID: 3041053
  40. Unwinding of duplex DNA from the SV40 origin of replication by T antigen.
    Science. 1987 Nov 13;238(4829):964-7 PMID: 2823389
  41. The unwinding of duplex regions in DNA by the simian virus 40 large tumor antigen-associated DNA helicase activity.
    J Biol Chem. 1988 Jan 5;263(1):383-92 PMID: 2826443
  42. Simian virus 40 large T antigen DNA helicase. Characterization of the ATPase-dependent DNA unwinding activity and its substrate requirements.
    J Biol Chem. 1988 Jan 5;263(1):436-42 PMID: 2826446
  43. Structure and function of SV40 large-T antigen.
    Philos Trans R Soc Lond B Biol Sci. 1987 Dec 15;317(1187):455-69 PMID: 2450379
  44. Geometry of the simian virus 40 large tumor antigen-DNA complex as probed by protease digestion.
    Proc Natl Acad Sci U S A. 1988 Apr;85(7):2086-90 PMID: 2832846
  45. Purification and characterization of replication protein A, a cellular protein required for in vitro replication of simian virus 40 DNA.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2523-7 PMID: 2833742
  46. Expression of simian virus 40 T antigen in Escherichia coli: localization of T-antigen origin DNA-binding domain to within 129 amino acids.
    J Virol. 1988 Jun;62(6):1999-2006 PMID: 2835505
  47. Simian virus 40 (SV40) T antigen binds specifically to double-stranded DNA but not to single-stranded DNA or DNA/RNA hybrids containing the SV40 regulatory sequences.
    J Virol. 1988 Jun;62(6):2204-8 PMID: 3367427
  48. Localized melting and structural changes in the SV40 origin of replication induced by T-antigen.
    EMBO J. 1988 Oct;7(10):3149-58 PMID: 2846276
  49. ATP-dependent assembly of double hexamers of SV40 T antigen at the viral origin of DNA replication.
    Nature. 1989 Apr 20;338(6217):658-62 PMID: 2539565
  50. Large T-antigen mutants define multiple steps in the initiation of simian virus 40 DNA replication.
    J Virol. 1989 Oct;63(10):4181-8 PMID: 2550664
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.72.12.10256-10259.1998
Published
1998-12-00
Pages
10256-9
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC110609
Subset
IM
Grants
NCI NIH HHS · R01 CA036118 · United States
NCI NIH HHS · CA36118 · United States
External Links
PubMed source DOI: 10.1128/JVI.72.12.10256-10259.1998 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: [email protected]

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