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PMID: 2152834 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Regulation of pp60c-src and its interaction with polyomavirus middle T antigen in insect cells.

Journal of virology ·Vol. 64 ·No. 1 ·1990-01-00 ·Pages 61-8

Piwnica-Worms H, Williams NG, Cheng SH, Roberts TM

Abstract

High yields of soluble, biologically active pp60c-src and middle t antigen (MTAg) of polyomavirus were produced in insect cells, using a baculovirus expression system. In mammalian cells, pp60c-src undergoes a regulatory phosphorylation on Tyr-527 in vivo and is autophosphorylated on Tyr-416 in vitro. In insect cells, pp60c-src was phosphorylated primarily on Tyr-416, although Tyr-527 was detectable at a low level. A kinase-negative mutant of pp60c-src was not phosphorylated on either Tyr-527 or Tyr-416 in insect cells and thus is an excellent biochemical reagent to search for the regulatory kinase that usually phosphorylates Tyr-527 in mammalian cells. MTAg synthesized in insect cells was not phosphorylated on tyrosine residues in vivo or in vitro, suggesting that it did not associate with any endogenous tyrosine kinases. However, MTAg isolated from cells coinfected with viruses encoding both MTAg and pp60c-src was phosphorylated on tyrosine residues both in vivo and in vitro.

MeSH Terms
Animals Antigens, Polyomavirus Transforming/biosynthesis,genetics Cell Line Gene Expression Regulation, Viral Genes, Viral Genetic Vectors Insect Viruses/genetics Insecta Methionine/metabolism Myristic Acid Myristic Acids/metabolism Oncogene Protein pp60(v-src)/biosynthesis,genetics Peptide Mapping Phosphopeptides/isolation & purification Phosphorylation Polyomavirus/genetics Protein Kinases/genetics,metabolism Viral Structural Proteins/genetics
Chemicals
Antigens, Polyomavirus Transforming Myristic Acids Phosphopeptides Viral Structural Proteins Myristic Acid Methionine Protein Kinases Oncogene Protein pp60(v-src)
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Piwnica-Worms H
Dana-Farber Cancer Institute, Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115.
Williams N G
Cheng S H
Roberts T M
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.64.1.61-68.1990
Published
1990-01-00
Pages
61-8
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC249044
Subset
IM
Grants
NCI NIH HHS · CA30002 · United States
NCI NIH HHS · CA43803 · United States
NCI NIH HHS · R29 CA50767 · United States
External Links
PubMed source DOI: 10.1128/JVI.64.1.61-68.1990 PMC Full Text View journal details
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