Home LiteratureArticle Details
PMID: 2987799 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Expression of polyoma early gene products in E. coli.

Nucleic acids research ·Vol. 13 ·No. 2 ·1985-01-25 ·Pages 501-19

Schaffhausen B, Benjamin TL, Lodge J, Kaplan D, Roberts TM

Abstract

The three products of the early region of polyoma virus have been cloned for expression in E. coli using the Tac promoter. Although the identical promoter and ribosome binding site are used in each final construction, the observed level of protein expression is different for each protein. While plasmids expressing wild type T antigens as well as a plasmid expressing the truncated Py-1387T middle T antigen lacking the membrane-anchoring sequence give rise to synthesis of proteins readily detectible by 35S-methionine labeling and immunoprecipitation, only small T and the middle T of Py-1387T are made in amounts sufficient for ready detection in total cell protein. Unlike middle T expressed in animal cells, middle T produced in E. coli is not detectibly phosphorylated. Further, the E. coli protein lacks tyrosine kinase activity.

MeSH Terms
Antigens, Viral, Tumor/genetics Cloning, Molecular Escherichia coli/genetics Gene Expression Regulation Genes, Viral Molecular Weight Peptide Fragments/analysis Polyomavirus/genetics Protein Kinases/genetics Protein-Tyrosine Kinases Viral Proteins/genetics beta-Galactosidase/genetics
Chemicals
Antigens, Viral, Tumor Peptide Fragments Viral Proteins Protein Kinases Protein-Tyrosine Kinases beta-Galactosidase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Schaffhausen B
Benjamin T L
Lodge J
Kaplan D
Roberts T M
References (51)
51 references, click to expand
  1. Expression of the gene for the polyoma small T antigen in Escherichia coli.
    J Virol. 1980 Oct;36(1):125-32 PMID: 6255200
  2. CELL-TRANSFORMING ABILITY OF A TEMPERATURE-SENSITIVE MUTANT OF POLYOMA VIRUS.
    Proc Natl Acad Sci U S A. 1965 Mar;53:486-91 PMID: 14338226
  3. Polyoma large T antigen regulates the integration of viral DNA sequences into the genome of transformed cells.
    Cell. 1981 Feb;23(2):347-55 PMID: 6258800
  4. 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
  5. Comparison of phosphorylation of two polyoma virus middle T antigens in vivo and in vitro.
    J Virol. 1981 Oct;40(1):184-96 PMID: 6169848
  6. The product of the lon (capR) gene in Escherichia coli is the ATP-dependent protease, protease La.
    Proc Natl Acad Sci U S A. 1981 Aug;78(8):4931-5 PMID: 6458037
  7. Rous sarcoma virus transforming protein, p60src, expressed in E. coli, functions as a protein kinase.
    Nature. 1981 Dec 24;294(5843):771-3 PMID: 6172716
  8. Mutation causing premature termination of the polyoma virus medium T antigen blocks cell transformation.
    J Virol. 1982 Mar;41(3):1014-24 PMID: 6284958
  9. 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
  10. Construction of overproducers of the bacteriophage 434 repressor and cro proteins.
    J Mol Appl Genet. 1981;1(2):139-47 PMID: 6286819
  11. The structures of the spliced mRNAs encoding polyoma virus early region proteins.
    J Mol Appl Genet. 1981;1(2):83-92 PMID: 6286820
  12. Syntheses and stabilities of proteins related to the polyoma small T antigen in Escherichia coli.
    Mol Cell Biol. 1982 Jan;2(1):88-92 PMID: 6287223
  13. Expression of an Abelson murine leukemia virus-encoded protein in Escherichia coli causes extensive phosphorylation of tyrosine residues.
    J Biol Chem. 1982 Nov 25;257(22):13181-4 PMID: 6292177
  14. Polyoma virus middle T antigen: relationship to cell membranes and apparent lack of ATP-binding activity.
    Mol Cell Biol. 1982 Oct;2(10):1187-98 PMID: 6184609
  15. The roles of individual polyoma virus early proteins in oncogenic transformation.
    Nature. 1982 Dec 23;300(5894):713-8 PMID: 6294529
  16. Purification of biologically active simian virus 40 small tumor antigen.
    Proc Natl Acad Sci U S A. 1983 Feb;80(4):906-10 PMID: 6302673
  17. 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
  18. Host range transforming gene of polyoma virus plays a role in virus assembly.
    Proc Natl Acad Sci U S A. 1983 Jun;80(12):3613-7 PMID: 6304724
  19. Transforming activity of polyoma virus middle-T antigen probed by site-directed mutagenesis.
    Nature. 1983 Aug 4-10;304(5925):456-9 PMID: 6308461
  20. Expression of the large T protein of polyoma virus promotes the establishment in culture of "normal" rodent fibroblast cell lines.
    Proc Natl Acad Sci U S A. 1983 Jul;80(14):4354-8 PMID: 6308618
  21. Role of the three polyoma virus early proteins in tumorigenesis.
    Mol Cell Biol. 1983 Aug;3(8):1451-9 PMID: 6312293
  22. Expression of the PDGF-related transforming protein of simian sarcoma virus in E. coli.
    Cell. 1984 Jan;36(1):43-9 PMID: 6319011
  23. 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
  24. Polyoma middle-sized T antigen can be phosphorylated on tyrosine at multiple sites in vitro.
    EMBO J. 1984 Jan;3(1):73-9 PMID: 6200324
  25. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  26. Mutants of Escherichia coli with a defect in the degradation of nonsense fragments.
    Nat New Biol. 1973 Jun 20;243(129):238-41 PMID: 4577438
  27. Polyoma gene function required for viral DNA synthesis.
    Virology. 1973 Sep;55(1):127-35 PMID: 4353950
  28. Localization of gene functions in polyoma virus DNA.
    Proc Natl Acad Sci U S A. 1976 Nov;73(11):4169-73 PMID: 186787
  29. Construction of plasmids carrying the cI gene of bacteriophage lambda.
    Proc Natl Acad Sci U S A. 1976 Nov;73(11):4174-8 PMID: 1069307
  30. Purification of simian virus 40 tumor antigen from a line of simian virus 40-transformed human cells.
    Proc Natl Acad Sci U S A. 1977 Sep;74(9):3745-9 PMID: 198802
  31. The binding site on SV40 DNA for a T antigen-related protein.
    Cell. 1978 Jan;13(1):165-79 PMID: 202398
  32. Tumor antigen(s) in cell productively infected by wild-type polyoma virus and mutant NG-18.
    Proc Natl Acad Sci U S A. 1978 Jan;75(1):79-83 PMID: 203944
  33. Virus-specific early RNA in 3T6 cells infected by a tsA mutant of polyoma virus.
    Virology. 1978 Mar;85(1):222-30 PMID: 206004
  34. Phenotypic expression in E. coli of a DNA sequence coding for mouse dihydrofolate reductase.
    Nature. 1978 Oct 19;275(5681):617-24 PMID: 360074
  35. A general method for maximizing the expression of a cloned gene.
    Proc Natl Acad Sci U S A. 1979 Feb;76(2):760-4 PMID: 370836
  36. Complete nucleotide sequence of the Escherichia coli plasmid pBR322.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:77-90 PMID: 383387
  37. Polyoma virus-specific 55K protein isolated from plasma membrane of productively infected cells is virus-coded and important for cell transformation.
    Virology. 1979 Oct 15;98(1):261-6 PMID: 225866
  38. Cloning of the active thymidine kinase gene of herpes simplex virus type 1 in Escherichia coli K-12.
    Proc Natl Acad Sci U S A. 1979 Aug;76(8):3755-9 PMID: 226978
  39. Construction and analysis of viable deletion mutants of polyoma virus.
    J Virol. 1979 Nov;32(2):523-9 PMID: 228075
  40. Phosphorylation of polyoma T antigens.
    Cell. 1979 Dec;18(4):935-46 PMID: 229974
  41. Synthesis of simian virus 40 t antigen in Escherichia coli.
    Proc Natl Acad Sci U S A. 1979 Nov;76(11):5596-600 PMID: 230483
  42. Viable deletion mutant in the medium and large T-antigen-coding sequences of the polyoma virus genome.
    J Virol. 1980 Mar;33(3):1215-20 PMID: 6245272
  43. Maximizing gene expression on a plasmid using recombination in vitro.
    Methods Enzymol. 1979;68:473-82 PMID: 232222
  44. Improved methods for maximizing expression of a cloned gene: a bacterium that synthesizes rabbit beta-globin.
    Cell. 1980 Jun;20(2):543-53 PMID: 6248249
  45. Middle T antigen as primary inducer of full expression of the phenotype of transformation by polyoma virus.
    J Virol. 1980 Jul;35(1):219-32 PMID: 6251270
  46. SV40 17K protein is associated with two cellular proteins.
    Cold Spring Harb Symp Quant Biol. 1980;44 Pt 1,:211-4 PMID: 6253134
  47. Expression of the human fibroblast interferon gene in Escherichia coli.
    Proc Natl Acad Sci U S A. 1980 Sep;77(9):5230-3 PMID: 6159642
  48. An antibody to a synthetic peptide recognizes polyomavirus middle-T antigen and reveals multiple in vitro tyrosine phosphorylation sites.
    Mol Cell Biol. 1984 Jul;4(7):1334-42 PMID: 6095063
  49. High-level expression of oncogenes in Escherichia coli.
    Gene Amplif Anal. 1983;3:147-74 PMID: 6101025
  50. Purification of polyoma virus.
    Virology. 1963 Feb;19:158-68 PMID: 14001211
  51. The tumor antigens and the early functions of polyoma virus.
    Mol Cell Biochem. 1980 Sep 15;32(2):63-93 PMID: 6256621
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/13.2.501
Published
1985-01-25
Pages
501-19
Language
English
Region
England
NLM ID
0411011
PMCID
PMC341011
Subset
IM
Grants
NCI NIH HHS · R01 CA034722 · United States
NCI NIH HHS · R01-CA19567 · United States
NCI NIH HHS · R01-CA30002 · United States
NCI NIH HHS · R01-CA34722 · United States
External Links
PubMed source DOI: 10.1093/nar/13.2.501 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