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PMID: 3011402 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mutations around the NG59 lesion indicate an active association of polyoma virus middle-T antigen with pp60c-src is required for cell transformation.

The EMBO journal ·Vol. 5 ·No. 2 ·1986-02-00 ·Pages 325-34

Cheng SH, Markland W, Markham AF, Smith AE

Abstract

The transforming activity of polyoma virus middle-T antigen is believed to be dependent on its ability to form a complex with the cellular tyrosine protein kinase, pp60c-src. This hypothesis is based on observations of mutants of middle-T which demonstrated a correlation between these two activities. To investigate further the significance of pp60c-src association in transformation by middle-T, a series of deletion and point mutants were constructed around the NG59 lesion since this region has been implicated in pp60c-src binding. Analysis of the middle-T variants revealed a complete correlation between the presence of associated activated pp60c-src and the ability to transform. Further, this ability of pp60c-src to associate with middle-T may depend on the presence of a beta-turn between amino acids 177 and 180. The results indicate the NG59 phenotype results from the introduction of an isoleucine residue between amino acids 177 and 178 rather than the transition mutation at 179. The mutant MG1 is a single point mutation (at residue 180) and represents the smallest change in the middle-T which abolishes both the transformating and kinase activity of middle-T. Taken together, the data suggest the region surrounding the NG59 lesion is involved in the formation of an active complex between middle-T and pp60c-src and strongly suggest that this association is an absolute requirement for polyoma virus-induced transformation.

MeSH Terms
Amino Acid Sequence Animals Antigens, Polyomavirus Transforming Antigens, Viral, Tumor/genetics Base Sequence Cell Line Cell Transformation, Neoplastic Chromosome Deletion DNA Restriction Enzymes DNA Transposable Elements Mutation Oncogene Protein pp60(v-src) Oncogene Proteins, Viral/genetics Plasmids Polyomavirus/genetics Protein Kinases/genetics Rats Retroviridae Proteins/genetics
Chemicals
Antigens, Polyomavirus Transforming Antigens, Viral, Tumor DNA Transposable Elements Oncogene Proteins, Viral Retroviridae Proteins Protein Kinases Oncogene Protein pp60(v-src) DNA Restriction Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Cheng S H
Markland W
Markham A F
Smith A E
References (34)
34 references, click to expand
  1. N-terminal amino acid sequences of the polyoma middle-size T antigen are important for protein kinase activity and cell transformation.
    Mol Cell Biol. 1984 May;4(5):817-21 PMID: 6328268
  2. The complex of polyoma virus middle-T antigen and pp60c-src.
    EMBO J. 1984 Mar;3(3):585-91 PMID: 6325177
  3. Enhancement of cellular src gene product associated tyrosyl kinase activity following polyoma virus infection and transformation.
    Cell. 1984 Oct;38(3):767-77 PMID: 6091900
  4. In vitro mutagenesis of a putative DNA binding domain of SV40 large-T.
    Virology. 1984 Nov;139(1):109-37 PMID: 6093377
  5. Middle tumor antigen of polyomavirus transformation-defective mutant NG59 is associated with pp60c-src.
    J Virol. 1985 Jan;53(1):114-9 PMID: 2981329
  6. A new member of the polyomavirus family: the hamster papovavirus. Complete nucleotide sequence and transformation properties.
    EMBO J. 1985 May;4(5):1279-86 PMID: 2988942
  7. Tyrosine phosphorylation within the amino-terminal domain of pp60c-src molecules associated with polyoma virus middle-sized tumor antigen.
    Proc Natl Acad Sci U S A. 1985 Jul;82(14):4568-72 PMID: 3927285
  8. Activation of the pp60c-src kinase by middle T antigen binding or by dephosphorylation.
    EMBO J. 1985 Jun;4(6):1471-7 PMID: 2411538
  9. Regulation of cellular phenotype and expression of polyomavirus middle T antigen in rat fibroblasts.
    Mol Cell Biol. 1985 Sep;5(9):2476-86 PMID: 2426583
  10. AGAR SUSPENSION CULTURE FOR THE SELECTIVE ASSAY OF CELLS TRANSFORMED BY POLYOMA VIRUS.
    Virology. 1964 Jun;23:291-4 PMID: 14187925
  11. Expression of polyoma early gene products in E. coli.
    Nucleic Acids Res. 1985 Jan 25;13(2):501-19 PMID: 2987799
  12. The reverse turn as a polypeptide conformation in globular proteins.
    Proc Natl Acad Sci U S A. 1973 Feb;70(2):538-42 PMID: 4510294
  13. A new technique for the assay of infectivity of human adenovirus 5 DNA.
    Virology. 1973 Apr;52(2):456-67 PMID: 4705382
  14. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene.
    Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961-5 PMID: 1105573
  15. Empirical predictions of protein conformation.
    Annu Rev Biochem. 1978;47:251-76 PMID: 354496
  16. Hybridization of synthetic oligodeoxyribonucleotides to phi chi 174 DNA: the effect of single base pair mismatch.
    Nucleic Acids Res. 1979 Aug 10;6(11):3543-57 PMID: 158748
  17. Identification of DNA sequence changes leading to loss of transforming ability in polyoma virus.
    J Biol Chem. 1980 Jan 10;255(1):230-5 PMID: 6243123
  18. Protein kinase activity associated with polyoma virus middle T antigen in vitro.
    Cell. 1979 Dec;18(4):915-24 PMID: 229972
  19. An activity phosphorylating tyrosine in polyoma T antigen immunoprecipitates.
    Cell. 1979 Dec;18(4):925-33 PMID: 229973
  20. Phosphorylation of polyoma T antigens.
    Cell. 1979 Dec;18(4):935-46 PMID: 229974
  21. DNA sequence analysis by primed synthesis.
    Methods Enzymol. 1980;65(1):560-80 PMID: 6154869
  22. Evidence that the phosphorylation of tyrosine is essential for cellular transformation by Rous sarcoma virus.
    Cell. 1980 Jul;20(3):807-16 PMID: 6251974
  23. Is polyoma virus middle T antigen a protein kinase?
    Cold Spring Harb Symp Quant Biol. 1980;44 Pt 1,:141-7 PMID: 6253127
  24. Transformation of rat cells by an altered polyoma virus genome expressing only the middle-T protein.
    Nature. 1981 Aug 13;292(5824):595-600 PMID: 6265803
  25. Total synthesis of a human leukocyte interferon gene.
    Nature. 1981 Aug 20;292(5825):756-62 PMID: 6167861
  26. Monoclonal antibodies against polyoma virus tumor antigens.
    Proc Natl Acad Sci U S A. 1982 Feb;79(4):1059-63 PMID: 6175960
  27. Carboxy terminus of polyoma middle-sized tumor antigen is required for attachment to membranes, associated protein kinase activities, and cell transformation.
    Proc Natl Acad Sci U S A. 1982 Jun;79(11):3579-83 PMID: 6179082
  28. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.
    J Mol Appl Genet. 1982;1(4):327-41 PMID: 6286831
  29. Deletion loop mutagenesis: a novel method for the construction of point mutations using deletion mutants.
    Nucleic Acids Res. 1982 Sep 11;10(17):5161-71 PMID: 6292832
  30. Differential subcellular localization of in vivo-phosphorylated and nonphosphorylated middle-sized tumor antigen of polyoma virus and its relationship to middle-sized tumor antigen phosphorylating activity in vitro.
    Proc Natl Acad Sci U S A. 1982 Nov;79(22):6812-6 PMID: 6294653
  31. Polyoma virus transforming protein associates with the product of the c-src cellular gene.
    Nature. 1983 Jun 2-8;303(5916):435-9 PMID: 6304524
  32. Transforming activity of polyoma virus middle-T antigen probed by site-directed mutagenesis.
    Nature. 1983 Aug 4-10;304(5925):456-9 PMID: 6308461
  33. Construction and expression of a recombinant DNA gene encoding a polyomavirus middle-size tumor antigen with the carboxyl terminus of the vesicular stomatitis virus glycoprotein G.
    Mol Cell Biol. 1984 Feb;4(2):282-9 PMID: 6321957
  34. Sequence requirements for nuclear location of simian virus 40 large-T antigen.
    Nature. 1984 Sep 6-11;311(5981):33-8 PMID: 6088992
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1986-02-00
Pages
325-34
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1166736
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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