Home LiteratureArticle Details
PMID: 8139575 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

A unique subpopulation of murine DNA polymerase alpha/primase specifically interacts with polyomavirus T antigen and stimulates DNA replication.

Molecular and cellular biology ·Vol. 14 ·No. 4 ·1994-04-00 ·Pages 2767-76

Moses K, Prives C

Abstract

Murine cells or cell extracts support the replication of plasmids containing the replication origin (ori-DNA) of polyomavirus (Py) but not that of simian virus 40 (SV40), whereas human cells or cell extracts support the replication of SV40 ori-DNA but not that of Py ori-DNA. It was shown previously that fractions containing DNA polymerase alpha/primase from permissive cells allow viral ori-DNA replication to proceed in extracts of nonpermissive cells. To extend these observations, the binding of Py T antigen to both the permissive and nonpermissive DNA polymerase alpha/primase was examined. Py T antigen was retained by a murine DNA polymerase alpha/primase but not by a human DNA polymerase alpha/primase affinity column. Likewise, a Py T antigen affinity column retained DNA polymerase alpha/primase activity from murine cells but not from human cells. The murine fraction which bound to the Py T antigen column was able to stimulate Py ori-DNA replication in the nonpermissive extract. However, the DNA polymerase alpha/primase activity in this murine fraction constituted only a relatively small proportion (approximately 20 to 40%) of the total murine DNA polymerase alpha/primase that had been applied to the column. The DNA polymerase alpha/primase purified from the nonbound murine fraction, although far more replete in this activity, was incapable of supporting Py DNA replication. The two forms of murine DNA polymerase alpha/primase also differed in their interactions with Py T antigen. Our data thus demonstrate that there are two distinct populations of DNA polymerase alpha/primase in murine cells and that species-specific interactions between T antigen and DNA polymerases can be identified. They may also provide the basis for initiating a novel means of characterizing unique subpopulations of DNA polymerase alpha/primase.

MeSH Terms
Animals Antigens, Polyomavirus Transforming/isolation & purification,metabolism Cell Line Chromatography, Affinity DNA Primase DNA Replication DNA, Viral/biosynthesis DNA-Directed DNA Polymerase/isolation & purification,metabolism HeLa Cells Humans Kinetics Mice Plasmids/biosynthesis Polyomavirus/genetics Protein Binding RNA Nucleotidyltransferases/isolation & purification,metabolism Simian virus 40/genetics Species Specificity
Chemicals
Antigens, Polyomavirus Transforming DNA, Viral DNA Primase RNA Nucleotidyltransferases DNA-Directed DNA Polymerase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Moses K
Department of Biological Sciences, Columbia University, New York, New York 10027.
Prives C
References (55)
55 references, click to expand
  1. Species-specific in vitro synthesis of DNA containing the polyoma virus origin of replication.
    Proc Natl Acad Sci U S A. 1986 Sep;83(17):6347-51 PMID: 3018726
  2. The role of the 70 kDa subunit of human DNA polymerase alpha in DNA replication.
    EMBO J. 1993 Dec;12(12):4555-66 PMID: 8223465
  3. Polyomavirus small t antigen: overproduction in bacteria, purification, and utilization for monoclonal and polyclonal antibody production.
    J Virol. 1986 Dec;60(3):1075-84 PMID: 3023660
  4. T-antigen-DNA polymerase alpha complex implicated in simian virus 40 DNA replication.
    Mol Cell Biol. 1986 Nov;6(11):4077-87 PMID: 3025630
  5. 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
  6. Expression of polyomavirus large T antigen by using a baculovirus vector.
    J Virol. 1987 May;61(5):1712-6 PMID: 3033300
  7. 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
  8. p53 and DNA polymerase alpha compete for binding to SV40 T antigen.
    Nature. 1987 Oct 1-7;329(6138):456-8 PMID: 3309672
  9. Immunoaffinity-purified DNA polymerase alpha displays novel properties.
    Biochemistry. 1987 Dec 15;26(25):8458-66 PMID: 3442669
  10. 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
  11. Expression and complex formation of simian virus 40 large T antigen and mouse p53 in insect cells.
    J Virol. 1988 Sep;62(9):3109-19 PMID: 2457111
  12. Recognition mechanisms in the synthesis of animal virus DNA.
    Biochem J. 1989 Feb 15;258(1):3-16 PMID: 2649089
  13. 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
  14. The murine p53 protein blocks replication of SV40 DNA in vitro by inhibiting the initiation functions of SV40 large T antigen.
    Cell. 1989 May 5;57(3):379-92 PMID: 2541911
  15. DNA primase-DNA polymerase alpha assembly from mouse FM3A cells. Purification of constituting enzymes, reconstitution, and analysis of RNA priming as coupled to DNA synthesis.
    J Biol Chem. 1989 Jun 15;264(17):10065-71 PMID: 2722861
  16. Initiation of simian virus 40 DNA replication in vitro: pulse-chase experiments identify the first labeled species as topologically unwound.
    Proc Natl Acad Sci U S A. 1989 Jun;86(11):3944-8 PMID: 2542958
  17. Animal virus DNA replication.
    Annu Rev Biochem. 1989;58:671-717 PMID: 2549858
  18. Initiation of eukaryotic DNA replication in vitro.
    Annu Rev Cell Biol. 1989;5:197-245 PMID: 2557059
  19. Binding and unwinding--how T antigen engages the SV40 origin of DNA replication.
    Cell. 1990 Jan 26;60(2):181-4 PMID: 2153460
  20. The role of human single-stranded DNA binding protein and its individual subunits in simian virus 40 DNA replication.
    J Biol Chem. 1990 May 5;265(13):7693-700 PMID: 2159011
  21. DNA helicase and nucleoside-5'-triphosphatase activities of polyoma virus large tumor antigen.
    Biochemistry. 1990 Jan 30;29(4):1003-9 PMID: 2160269
  22. ATP phosphohydrolase (ATPase) activity of a polyoma virus T antigen.
    Eur J Biochem. 1980 Aug;109(2):553-60 PMID: 6250847
  23. 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
  24. Preparation and preliminary characterization of monoclonal antibodies against human DNA polymerase alpha.
    J Biol Chem. 1982 Jul 25;257(14):8386-90 PMID: 7085672
  25. Polyomavirus and simian virus 40 large T antigens bind to common DNA sequences.
    J Virol. 1984 Mar;49(3):925-37 PMID: 6321773
  26. Simian virus 40 DNA replication in vitro.
    Proc Natl Acad Sci U S A. 1984 Nov;81(22):6973-7 PMID: 6095264
  27. Middle tumor antigen of polyomavirus transformation-defective mutant NG59 is associated with pp60c-src.
    J Virol. 1985 Jan;53(1):114-9 PMID: 2981329
  28. Simian virus 40 and polyomavirus large tumor antigens have different requirements for high-affinity sequence-specific DNA binding.
    J Virol. 1985 May;54(2):532-45 PMID: 2985816
  29. Simian virus 40 DNA replication in vitro: specificity of initiation and evidence for bidirectional replication.
    Mol Cell Biol. 1985 Jun;5(6):1238-46 PMID: 2993858
  30. Differential inhibitors of DNA polymerases alpha and delta.
    Biochem Biophys Res Commun. 1985 Oct 30;132(2):628-34 PMID: 4062942
  31. Guanine nucleotide contacts within viral DNA sequences bound by polyomavirus large T antigen.
    J Virol. 1986 Feb;57(2):505-14 PMID: 3003383
  32. Role of DNA polymerase alpha and DNA primase in simian virus 40 DNA replication in vitro.
    Proc Natl Acad Sci U S A. 1986 May;83(9):2869-73 PMID: 3010320
  33. Common regulatory elements control gene expression from polyoma early and late promoters in cells transformed by chimeric plasmids.
    Mol Cell Biol. 1985 Aug;5(8):2070-9 PMID: 3018549
  34. The replication functions of SV40 T antigen are regulated by phosphorylation.
    Cell. 1990 Jun 1;61(5):735-8 PMID: 2160857
  35. Sequential initiation of lagging and leading strand synthesis by two different polymerase complexes at the SV40 DNA replication origin.
    Nature. 1990 Aug 9;346(6284):534-9 PMID: 2165567
  36. A third essential DNA polymerase in S. cerevisiae.
    Cell. 1990 Sep 21;62(6):1143-51 PMID: 2169349
  37. SV40 T antigen binds directly to the large subunit of purified DNA polymerase alpha.
    EMBO J. 1990 Oct;9(10):3329-36 PMID: 1698613
  38. Reconstitution of simian virus 40 DNA replication with purified proteins.
    Proc Natl Acad Sci U S A. 1990 Nov;87(22):8692-6 PMID: 2174160
  39. The DNA-binding properties of polyomavirus large T antigen are altered by ATP and other nucleotides.
    J Virol. 1991 Feb;65(2):687-99 PMID: 1846192
  40. Replication factors required for SV40 DNA replication in vitro. I. DNA structure-specific recognition of a primer-template junction by eukaryotic DNA polymerases and their accessory proteins.
    J Biol Chem. 1991 Jan 25;266(3):1950-60 PMID: 1671045
  41. Replication factors required for SV40 DNA replication in vitro. II. Switching of DNA polymerase alpha and delta during initiation of leading and lagging strand synthesis.
    J Biol Chem. 1991 Jan 25;266(3):1961-8 PMID: 1671046
  42. Properties of DNA polymerases delta and epsilon, and their roles in eukaryotic DNA replication.
    Biochim Biophys Acta. 1991 Jan 17;1088(1):11-24 PMID: 1846563
  43. 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
  44. Cell cycle-dependent phosphorylation of human DNA polymerase alpha.
    J Biol Chem. 1991 Apr 25;266(12):7893-903 PMID: 1902230
  45. Characterization of a cDNA encoding the 70-kDa single-stranded DNA-binding subunit of human replication protein A and the role of the protein in DNA replication.
    J Biol Chem. 1991 Jun 25;266(18):12090-8 PMID: 2050703
  46. DNA helicase and duplex DNA fragment unwinding activities of polyoma and simian virus 40 large T antigen display similarities and differences.
    J Biol Chem. 1991 Jul 5;266(19):12668-75 PMID: 1648101
  47. Mouse DNA primase plays the principal role in determination of permissiveness for polyomavirus DNA replication.
    J Virol. 1991 Sep;65(9):4874-81 PMID: 1651410
  48. ATP induces the assembly of polyoma large tumor antigen into hexamers.
    Virology. 1991 Sep;184(1):399-403 PMID: 1714665
  49. Eukaryotic DNA polymerases.
    Annu Rev Biochem. 1991;60:513-52 PMID: 1883203
  50. 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
  51. Structure and function of simian virus 40 large tumor antigen.
    Annu Rev Biochem. 1992;61:55-85 PMID: 1323237
  52. 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
  53. 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
  54. Purification and functional characterization of bovine RP-A in an in vitro SV40 DNA replication system.
    Chromosoma. 1992;102(1 Suppl):S52-9 PMID: 1337880
  55. DNA helicase activity of SV40 large tumor antigen.
    EMBO J. 1986 Aug;5(8):1939-44 PMID: 3019672
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.4.2767-2776.1994
Published
1994-04-00
Pages
2767-76
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358642
Subset
IM
Grants
NCI NIH HHS · CA-26905 · United States
External Links
PubMed source DOI: 10.1128/mcb.14.4.2767-2776.1994 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