In vitro phosphorylation of SV40 large T antigen.

Phosphorylation of simian virus 40 large T antigen (large T) was investigated in vitro. "Autophosphorylation" of large T resulted in the modification of Ser106, Ser112, Ser123, Thr124, either Ser676, Ser677, or Ser679, and Thr701. All of these residues were also found to be phosphorylated in vivo. Reaction of large T with purified casein kinase I resulted in phosphorylation of Ser123, possibly Thr124, and either Ser676, Ser677, or Ser679, while purified casein kinase II phosphorylated Ser106 and possibly Ser112. Submolar amounts of phosphate were transferred to large T indicating that only a fraction of large T served as substrate for the casein kinases. Removal of serine-bound phosphate did not affect the subsequent autophosphorylation or phosphorylation by casein kinase I and II. No phosphorylation at in vivo sites was observed with the cAMP-, cGMP-, or Ca2+/phospholipid-dependent protein kinases, or with the protease-activated kinase I and II.

[1]  D. Kalderon,et al.  In vitro mutagenesis of a putative DNA binding domain of SV40 large-T. , 1984, Virology.

[2]  B. Stillman,et al.  Regulation of SV40 DNA replication by phosphorylation of T antigen. , 1987, The EMBO journal.

[3]  S. Tahara,et al.  Cyclic Nucleotide-independent protein kinases from rabbit reticulocytes. Identification and characterization of a protein kinase activated by proteolysis. , 1981, The Journal of biological chemistry.

[4]  M. Greaves,et al.  A one-step purification of membrane proteins using a high efficiency immunomatrix. , 1982, The Journal of biological chemistry.

[5]  K. Scheidtmann.  Phosphorylation of simian virus 40 large T antigen: cytoplasmic and nuclear phophorylation sites differ in their metabolic stability. , 1986, Virology.

[6]  C. Vesco,et al.  Deletion of 43 amino acids in the NH2-terminal half of the large tumor antigen of simian virus 40 results in a non-karyophilic protein capable of transforming established cells. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[7]  W. Fiers,et al.  Improved localization of phosphorylation sites in simian virus 40 large T antigen , 1983, Journal of virology.

[8]  G. Walter,et al.  Removal of serine phosphates from simian virus 40 large T antigen increases its ability to stimulate DNA replication in vitro but has no effect on ATPase and DNA binding , 1987, Journal of virology.

[9]  P. Tegtmeyer,et al.  Modification of simian virus 40 protein A , 1977, Journal of virology.

[10]  E. Krebs,et al.  Protein phosphorylation catalyzed by cyclic AMP-dependent and cyclic GMP-dependent protein kinases. , 1980, Annual review of pharmacology and toxicology.

[11]  R. Knippers,et al.  DNA helicase activity of SV40 large tumor antigen. , 1986, The EMBO journal.

[12]  H. Zuber,et al.  Amino acid sequence analysis of fragments generated by partial proteolysis from large simian virus 40 tumor antigen. , 1980, The Journal of biological chemistry.

[13]  R. Tjian.  The binding site on SV40 DNA for a T antigen-related protein , 1978, Cell.

[14]  G M Hathaway,et al.  Cyclic nucleotide-independent protein kinases from rabbit reticulocytes. Purification of casein kinases. , 1979, The Journal of biological chemistry.

[15]  F. Graham,et al.  Characteristics of a human cell line transformed by DNA from human adenovirus type 5. , 1977, The Journal of general virology.

[16]  R. Tjian.  Regulation of viral transcription and DNA replication by the SV40 large T antigen. , 1981, Current topics in microbiology and immunology.

[17]  G. Walter,et al.  Monoclonal antibodies specific for the carboxy terminus of simian virus 40 large T antigen , 1984, Journal of virology.

[18]  T. Lubben,et al.  Cyclic nucleotide-independent protein kinases from rabbit reticulocytes. Purification and characterization of protease-activated kinase II. , 1983, The Journal of biological chemistry.

[19]  R. Tjian,et al.  Catalytic properties of the SV40 large T antigen. , 1980, Cold Spring Harbor symposia on quantitative biology.

[20]  A. Smith,et al.  Characterization of the amino-terminal tryptic peptide of simian virus 40 small-t and large-T antigens , 1978, Journal of virology.

[21]  P. Tegtmeyer,et al.  Binding of dephosphorylated A protein to SV40 DNA. , 1981, Virology.

[22]  R. Tjian,et al.  T-antigen-DNA polymerase alpha complex implicated in simian virus 40 DNA replication , 1986, Molecular and cellular biology.

[23]  K. Scheidtmann,et al.  DNA-binding activity of simian virus 40 large T antigen correlates with a distinct phosphorylation state , 1984, Journal of virology.

[24]  Y. Gluzman.  Eukaryotic viral vectors , 1982 .

[25]  R. Tjian,et al.  Positive and negative regulation of transcription in vitro: Enhancer-binding protein AP-2 is inhibited by SV40 T antigen , 1987, Cell.

[26]  D. Simmons,et al.  Phosphorylation downregulates the DNA-binding activity of simian virus 40 T antigen , 1986, Journal of virology.

[27]  D. Lane,et al.  T antigen is bound to a host protein in SY40-transformed cells , 1979, Nature.

[28]  K. Scheidtmann,et al.  Complexes of polyoma virus medium T antigen and cellular proteins. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[29]  M. DePamphilis,et al.  Replication of SV40 and Polyoma Virus Chromosomes , 1986 .

[30]  D. Livingston,et al.  Protein kinase activity associated with simian virus 40 T antigen. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[31]  Butel Js.  SV40 large T-antigen: dual oncogene. , 1986 .

[32]  E. Baumann.  DNA-binding properties of phosphorylated and dephosphorylated D2-T antigen, a simian-virus-40 T-antigen-related protein. , 2008, European journal of biochemistry.

[33]  G. Hathaway,et al.  Casein kinases--multipotential protein kinases. , 1982, Current topics in cellular regulation.

[34]  R. Tjian,et al.  Biological activity of purified simian virus 40 T antigen proteins. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[35]  C. Merril,et al.  Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. , 1981, Science.