Home LiteratureArticle Details
PMID: 12634843 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Bidirectional DNA unwinding by a ternary complex of T antigen, nucleolin and topoisomerase I.

EMBO reports ·Vol. 4 ·No. 3 ·2003-03-00 ·Pages 263-8

Seinsoth S, Uhlmann-Schiffler H, Stahl H

Abstract

The simian virus 40 large tumour-antigen (T antigen) DNA helicase is a hexameric structure; it has been proposed that, in viral DNA replication, two of these hexamers are combined to form a bipartite holoenzyme that acts concurrently at both forks of a replication bubble. In a search for structural components of this helicase complex, we have identified nucleolin as a specific binding protein for the T-antigen hexamer. We show that nucleolin, in co-operation with human topoisomerase I, mediates the cohesion of the T-antigen helicase holoenzyme during plasmid unwinding. Our results provide biochemical evidence for a direct role of nucleolin in DNA replication, in addition to its known function in ribosome biogenesis. The data presented here suggest that nucleolin enables the formation of a functional 'helicase-swivelase' complex at the replication fork.

MeSH Terms
Amino Acid Sequence Animals Antigens, Viral, Tumor/chemistry,genetics,metabolism COS Cells Cell Line Chlorocebus aethiops DNA/chemistry,genetics,ultrastructure DNA Helicases/metabolism DNA Topoisomerases, Type I/metabolism HeLa Cells Humans Molecular Sequence Data Nuclear Proteins/metabolism Peptide Fragments/immunology Phosphoproteins/metabolism RNA-Binding Proteins/metabolism Recombinant Proteins/metabolism Simian virus 40/immunology Spodoptera Transfection
Chemicals
Antigens, Viral, Tumor Nuclear Proteins Peptide Fragments Phosphoproteins RNA-Binding Proteins Recombinant Proteins nucleolin DNA DNA Helicases DNA Topoisomerases, Type I
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Seinsoth Stephanie
Medizinische Biochemie und Molekularbiologie, Universität des Saarlandes, Gebaude 45, D-66421 Homburg, Germany.
Uhlmann-Schiffler Heike
Stahl Hans
References (28)
28 references, click to expand
  1. Dimerization of simian virus 40 T-antigen hexamers activates T-antigen DNA helicase activity.
    J Virol. 1997 Nov;71(11):8766-73 PMID: 9343236
  2. The initiation of simian virus 40 DNA replication in vitro.
    Crit Rev Biochem Mol Biol. 1997;32(6):503-68 PMID: 9444478
  3. Two-dimensional electrophoresis reveals a nuclear matrix-associated nucleolin complex of basic isoelectric point.
    Electrophoresis. 1997 Dec;18(14):2645-53 PMID: 9527495
  4. Interaction between the N-terminus of human topoisomerase I and SV40 large T antigen.
    Nucleic Acids Res. 1998 Apr 1;26(7):1841-7 PMID: 9512561
  5. Structure and functions of nucleolin.
    J Cell Sci. 1999 Mar;112 ( Pt 6):761-72 PMID: 10036227
  6. The organization of replication and transcription.
    Science. 1999 Jun 11;284(5421):1790-5 PMID: 10364545
  7. Eukaryotic DNA replication: a model for a fixed double replisome.
    Trends Genet. 2000 Feb;16(2):88-92 PMID: 10652536
  8. Topoisomerase I associates specifically with simian virus 40 large-T-antigen double hexamer-origin complexes.
    J Virol. 2000 Jun;74(11):5224-32 PMID: 10799598
  9. Formation of a complex between nucleolin and replication protein A after cell stress prevents initiation of DNA replication.
    J Cell Biol. 2000 May 15;149(4):799-810 PMID: 10811822
  10. Is the MCM2-7 complex the eukaryotic DNA replication fork helicase?
    Curr Opin Genet Dev. 2001 Feb;11(1):64-70 PMID: 11163153
  11. Regulation of dna replication after heat shock by replication protein a-nucleolin interactions.
    J Biol Chem. 2001 Jun 8;276(23):20579-88 PMID: 11389152
  12. Computer-assisted identification of cell cycle-related genes: new targets for E2F transcription factors.
    J Mol Biol. 2001 May 25;309(1):99-120 PMID: 11491305
  13. Inhibition of DNA replication and induction of S phase cell cycle arrest by G-rich oligonucleotides.
    J Biol Chem. 2001 Nov 16;276(46):43221-30 PMID: 11555643
  14. SV40 large T antigen hexamer structure: domain organization and DNA-induced conformational changes.
    Curr Biol. 2002 Mar 19;12(6):472-6 PMID: 11909532
  15. Yph1p, an ORC-interacting protein: potential links between cell proliferation control, DNA replication, and ribosome biogenesis.
    Cell. 2002 Jun 28;109(7):835-48 PMID: 12110181
  16. Preformed hexamers of SV40 T antigen are active in RNA and origin-DNA unwinding.
    Nucleic Acids Res. 2002 Jul 15;30(14):3192-201 PMID: 12136101
  17. RNA unwinding activity of SV40 large T antigen.
    Cell. 1989 Jun 16;57(6):955-63 PMID: 2472217
  18. Cloning and sequencing of the human nucleolin cDNA.
    FEBS Lett. 1989 Jun 19;250(1):99-105 PMID: 2737305
  19. Simultaneous purification of DNA topoisomerase I and II from eukaryotic cells.
    Prep Biochem. 1989;19(1):37-48 PMID: 2544867
  20. Mitosis-specific phosphorylation of nucleolin by p34cdc2 protein kinase.
    Mol Cell Biol. 1990 Jul;10(7):3607-18 PMID: 2192260
  21. Structural topography of simian virus 40 DNA replication.
    J Virol. 1991 May;65(5):2578-88 PMID: 1850031
  22. A comprehensive study on the isolation and characterization of the HeLa S3 nuclear matrix.
    J Cell Sci. 1991 Mar;98 ( Pt 3):281-91 PMID: 1711534
  23. 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
  24. The amino terminus of mammalian nucleolin specifically recognizes SV40 T-antigen type nuclear localization sequences.
    Eur J Cell Biol. 1993 Oct;62(1):13-21 PMID: 8269971
  25. Nucleolin is a matrix attachment region DNA-binding protein that specifically recognizes a region with high base-unpairing potential.
    Mol Cell Biol. 1995 Jan;15(1):456-65 PMID: 7799955
  26. Unwinding of chromatin by the SV40 large T antigen DNA helicase.
    EMBO J. 1995 Jul 3;14(13):3215-25 PMID: 7621834
  27. Simian virus 40 large T antigen binds to topoisomerase I.
    Virology. 1996 Aug 15;222(2):365-74 PMID: 8806520
  28. Nucleolin is one component of the B cell-specific transcription factor and switch region binding protein, LR1.
    Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3605-10 PMID: 9108024
Article Info
Journal
EMBO reports
Abbr.
EMBO Rep
ISSN
1469-221X
DOI
10.1038/sj.embor.embor770
Published
2003-03-00
Pages
263-8
Language
English
Region
England
NLM ID
100963049
PMCID
PMC1315898
Subset
IM
External Links
PubMed source DOI: 10.1038/sj.embor.embor770 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