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PMID: 3357207 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mutations in the phosphorylation sites of simian virus 40 (SV40) T antigen alter its origin DNA-binding specificity for sites I or II and affect SV40 DNA replication activity.

Journal of virology ·Vol. 62 ·No. 5 ·1988-05-00 ·Pages 1598-605

Schneider J, Fanning E

Abstract

A series of mutants of simian virus 40 was constructed by oligonucleotide-directed mutagenesis to study the role of phosphorylation in the functions of large T antigen. Each of the previously mapped phosphorylated serine and threonine residues in large T antigen was replaced by an alanine or cysteine residue or, in one case, by glutamic acid. Mutant DNAs were assayed for plaque-forming activity, viral DNA replication, expression of T antigen, and morphological transformation of rat cells. Viable mutants were isolated, suggesting that modification of some residues is not essential for the biological functions of T antigen. Two of these mutants replicated more efficiently than did the wild type. Seven mutants were partially or completely deficient in viral DNA replication but retained cell transformation activity comparable with that of the wild-type protein. Biochemical analysis of the mutant T antigens demonstrated novel origin DNA-binding properties of several mutant proteins. The results are consistent with the idea that differential phosphorylation defines several functional subclasses of T-antigen molecules.

MeSH Terms
Animals Antigens, Polyomavirus Transforming/genetics Base Sequence Binding Sites Cell Transformation, Viral DNA Replication DNA, Viral/metabolism Mutation Phosphorylation Rats Transfection
Chemicals
Antigens, Polyomavirus Transforming DNA, Viral
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Schneider J
Institute for Biochemistry, Munich, Federal Republic of Germany.
Fanning E
References (66)
66 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. A new technique for the assay of infectivity of human adenovirus 5 DNA.
    Virology. 1973 Apr;52(2):456-67 PMID: 4705382
  3. Effect of cell chromosome number on simian virus 40 replication.
    Exp Cell Res. 1973 Sep;81(1):120-6 PMID: 4357029
  4. Detection of specific sequences among DNA fragments separated by gel electrophoresis.
    J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
  5. Modification of simian virus 40 protein A.
    J Virol. 1977 Feb;21(2):647-57 PMID: 189092
  6. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  7. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-sepharose.
    Immunochemistry. 1978 Jul;15(7):429-36 PMID: 30693
  8. Phosphorylation of T-antigen and control T-antigen expression in cells transformed by wild-type and tsA mutants of simian virus 40.
    J Virol. 1979 Feb;29(2):753-62 PMID: 219256
  9. Identification and characterization of fast-sedimenting SV40 nucleoprotein complexes.
    Virology. 1979 Jul 15;96(1):54-63 PMID: 223310
  10. Filter replicas and permanent collections of recombinant DNA plasmids.
    Nucleic Acids Res. 1979 Dec 20;7(8):2115-36 PMID: 118435
  11. SV40-transformed simian cells support the replication of early SV40 mutants.
    Cell. 1981 Jan;23(1):175-82 PMID: 6260373
  12. Detection and characterization of multiple forms of simian virus 40 large T antigen.
    J Virol. 1981 Jan;37(1):92-102 PMID: 6261004
  13. Complex of simian virus 40 large tumor antigen and 48,000-dalton host tumor antigen.
    Proc Natl Acad Sci U S A. 1981 Jan;78(1):105-9 PMID: 6941238
  14. Normal rat cell lines deficient in nuclear thymidine kinase.
    Virology. 1981 Aug;113(1):408-11 PMID: 7269249
  15. Monoclonal antibodies specific for simian virus 40 tumor antigens.
    J Virol. 1981 Sep;39(3):861-9 PMID: 6169844
  16. Binding of dephosphorylated A protein to SV40 DNA.
    Virology. 1981 Nov;115(1):88-96 PMID: 6270903
  17. The role of protein phosphorylation in neural and hormonal control of cellular activity.
    Nature. 1982 Apr 15;296(5858):613-20 PMID: 6280056
  18. Regulatory mutants of simian virus 40. Effect of mutations at a T antigen binding site on DNA replication and expression of viral genes.
    J Mol Biol. 1982 Apr 15;156(3):531-48 PMID: 6288959
  19. Phosphorylation pattern of large T antigens in mouse cells infected by simian virus 40 wild type or deletion mutants.
    J Virol. 1982 Sep;43(3):761-71 PMID: 6292459
  20. Simian virus 40 large T antigen is phosphorylated at multiple sites clustered in two separate regions.
    J Virol. 1982 Oct;44(1):116-33 PMID: 6292479
  21. Oligonucleotide-directed mutagenesis as a general and powerful method for studies of protein function.
    Proc Natl Acad Sci U S A. 1982 Nov;79(21):6409-13 PMID: 6983070
  22. Oligonucleotide-directed mutagenesis using M13-derived vectors: an efficient and general procedure for the production of point mutations in any fragment of DNA.
    Nucleic Acids Res. 1982 Oct 25;10(20):6487-500 PMID: 6757864
  23. Improved localization of phosphorylation sites in simian virus 40 large T antigen.
    J Virol. 1983 Jan;45(1):315-31 PMID: 6296439
  24. Metabolic turnover of phosphorylation sites in simian virus 40 large T antigen.
    J Virol. 1983 Jan;45(1):442-6 PMID: 6296451
  25. High efficiency polyoma DNA transfection of chloroquine treated cells.
    Nucleic Acids Res. 1983 Mar 11;11(5):1295-308 PMID: 6298741
  26. Mutational analysis of simian virus 40 T antigen: isolation and characterization of mutants with deletions in the T-antigen gene.
    Mol Cell Biol. 1983 Feb;3(2):203-13 PMID: 6300656
  27. Specific DNA binding activity of T antigen subclasses varies among different SV40-transformed cell lines.
    Virology. 1983 Apr 15;126(1):19-31 PMID: 6302981
  28. Improvements in the phosphoramidite procedure for the synthesis of oligodeoxyribonucleotides.
    Nucleic Acids Res. 1983 May 11;11(9):2575-84 PMID: 6856470
  29. Essential contact residues within SV40 large T antigen binding sites I and II identified by alkylation-interference.
    Cell. 1984 Jan;36(1):155-62 PMID: 6319005
  30. DNA-binding activity of simian virus 40 large T antigen correlates with a distinct phosphorylation state.
    J Virol. 1984 Apr;50(1):1-12 PMID: 6321781
  31. Transformation by polyoma virus is drastically reduced by substitution of phenylalanine for tyrosine at residue 315 of middle-sized tumor antigen.
    Proc Natl Acad Sci U S A. 1984 Feb;81(3):679-83 PMID: 6322163
  32. Differential phosphorylation of cytoplasmic and nuclear variants of simian virus 40 large T antigen encoded by simian virus 40-adenovirus 7 hybrid viruses.
    J Virol. 1984 May;50(2):636-40 PMID: 6323765
  33. Subclasses of simian virus 40 large T antigen: differential binding of two subclasses of T antigen from productively infected cells to viral and cellular DNA.
    EMBO J. 1982;1(9):1023-8 PMID: 6329720
  34. Stepwise phosphorylation of the NH2-terminal region of the simian virus 40 large T antigen.
    J Biol Chem. 1984 Jul 10;259(13):8633-40 PMID: 6330117
  35. Monoclonal antibody to simian virus 40 small t.
    J Virol. 1984 Sep;51(3):760-7 PMID: 6088798
  36. Sequence requirements for nuclear location of simian virus 40 large-T antigen.
    Nature. 1984 Sep 6-11;311(5981):33-8 PMID: 6088992
  37. In vitro mutagenesis of a putative DNA binding domain of SV40 large-T.
    Virology. 1984 Nov;139(1):109-37 PMID: 6093377
  38. Genetic and biochemical analysis of transformation-competent, replication-defective simian virus 40 large T antigen mutants.
    J Virol. 1985 Jan;53(1):120-7 PMID: 2981330
  39. Minimal size plasmids containing an M13 origin for production of single-strand transducing particles.
    J Mol Appl Genet. 1984;2(6):507-17 PMID: 6099398
  40. Mutations near the carboxyl terminus of the simian virus 40 large tumor antigen alter viral host range.
    J Virol. 1985 May;54(2):569-75 PMID: 2985819
  41. Two integrated partial repeats of simian virus 40 together code for a super-T antigen.
    Mol Cell Biol. 1985 Apr;5(4):742-50 PMID: 2985968
  42. Induction of early mitotic events in a cell-free system.
    Cell. 1985 May;41(1):165-75 PMID: 3888406
  43. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA.
    DNA. 1985 Apr;4(2):165-70 PMID: 3996185
  44. Site-directed mutagenesis of the simian virus 40 large T-antigen gene: replication-defective amino acid substitution mutants that retain the ability to induce morphological transformation.
    J Virol. 1985 Jul;55(1):1-9 PMID: 2989548
  45. Protein-tyrosine kinases.
    Annu Rev Biochem. 1985;54:897-930 PMID: 2992362
  46. Seventeen base pairs of region I encode a novel tripartite binding signal for SV40 T antigen.
    Cell. 1985 Sep;42(2):539-48 PMID: 2992803
  47. Identification and biochemical analysis of DNA replication-defective large T antigens from SV40-transformed cells.
    Virology. 1985 Oct 30;146(2):188-202 PMID: 2996217
  48. T antigen and template requirements for SV40 DNA replication in vitro.
    EMBO J. 1985 Nov;4(11):2933-9 PMID: 2998767
  49. Simian virus 40 origin DNA-binding domain on large T antigen.
    J Virol. 1986 Jan;57(1):50-64 PMID: 3001365
  50. Properties of the simian virus 40 (SV40) large T antigens encoded by SV40 mutants with deletions in gene A.
    J Virol. 1986 Feb;57(2):539-46 PMID: 3003386
  51. A cell free system to study reassembly of the nuclear envelope at the end of mitosis.
    Cell. 1986 Feb 28;44(4):639-52 PMID: 3948244
  52. Antigenic binding sites of monoclonal antibodies specific for simian virus 40 large T antigen.
    J Virol. 1986 Mar;57(3):1168-72 PMID: 2419584
  53. DNA-binding region of the simian virus 40 tumor antigen.
    J Virol. 1986 Mar;57(3):776-85 PMID: 3005626
  54. Phosphorylation of the human interleukin-2 receptor and a synthetic peptide identical to its C-terminal, cytoplasmic domain.
    J Biol Chem. 1986 Apr 15;261(11):5075-80 PMID: 3082877
  55. The cell cycle control gene cdc2+ of fission yeast encodes a protein kinase potentially regulated by phosphorylation.
    Cell. 1986 Apr 25;45(2):261-8 PMID: 3516412
  56. Replication and transformation functions of in vitro-generated simian virus 40 large T antigen mutants.
    J Virol. 1986 May;58(2):526-35 PMID: 3009866
  57. Allosteric control of simian virus 40 T-antigen binding to viral origin DNA.
    J Virol. 1986 Jun;58(3):765-72 PMID: 3009885
  58. Phosphorylation downregulates the DNA-binding activity of simian virus 40 T antigen.
    J Virol. 1986 Dec;60(3):888-94 PMID: 3023678
  59. trans-dominant defective mutants of simian virus 40 T antigen.
    J Virol. 1987 Feb;61(2):436-45 PMID: 3027373
  60. Activation and suppression of pp60c-src transforming ability by mutation of its primary sites of tyrosine phosphorylation.
    Cell. 1987 Apr 10;49(1):65-73 PMID: 3103925
  61. Tyrosine phosphorylation regulates the biochemical and biological properties of pp60c-src.
    Cell. 1987 Apr 10;49(1):75-82 PMID: 3103926
  62. Cell transformation by pp60c-src mutated in the carboxy-terminal regulatory domain.
    Cell. 1987 Apr 10;49(1):83-91 PMID: 3103927
  63. Regulation of SV40 DNA replication by phosphorylation of T antigen.
    EMBO J. 1987 Jan;6(1):153-60 PMID: 3034573
  64. Oligomerization and origin DNA-binding activity of simian virus 40 large T antigen.
    J Virol. 1987 Jul;61(7):2076-83 PMID: 3035209
  65. Selective extraction of polyoma DNA from infected mouse cell cultures.
    J Mol Biol. 1967 Jun 14;26(2):365-9 PMID: 4291934
  66. Enchancement of the infectivity of simian virus 40 deoxyribonucleic acid with diethylaminoethyl-dextran.
    J Natl Cancer Inst. 1968 Aug;41(2):351-7 PMID: 4299537
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.62.5.1598-1605.1988
Published
1988-05-00
Pages
1598-605
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC253187
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.62.5.1598-1605.1988 PMC Full Text View journal details
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