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PMID: 9420259 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Serine 257 phosphorylation regulates association of polyomavirus middle T antigen with 14-3-3 proteins.

Journal of virology ·Vol. 72 ·No. 1 ·1998-01-00 ·Pages 558-63

Culleré X, Rose P, Thathamangalam U, Chatterjee A, Mullane KP, Pallas DC, Benjamin TL, Roberts TM, Schaffhausen BS

Abstract

Polyomavirus middle T antigen (MT) is phosphorylated on serine residues. Partial proteolytic mapping and Edman degradation identified serine 257 as a major site of phosphorylation. This was confirmed by site-directed mutagenesis. Isoelectric focusing of immunoprecipitated MT from transfected 293T cells showed that phosphorylation on wild-type MT occurred at near molar stoichiometry at S257. MT was previously shown to be associated with 14-3-3 proteins, which have been connected to cell cycle regulation and signaling. The association of 14-3-3 proteins with MT depended on the serine 257 phosphorylation site. This has been demonstrated by comparing wild-type and S257A mutant MTs expressed with transfected 293T cells or with Sf9 cells infected with recombinant baculoviruses. The 257 site is not critical for transformation of fibroblasts in vitro, since S257A and S257C mutant MTs retained the ability to form foci or colonies in agar. The tumor profile of a virus expressing S257C MT showed a striking deficiency in the induction of salivary gland tumors. The basis for this defect is uncertain. However, differences in activity for the wild type and mutant MT lacking the 14-3-3 binding site have been observed in transient reporter assays.

MeSH Terms
14-3-3 Proteins 3T3 Cells Animals Antigens, Polyomavirus Transforming/chemistry,genetics,metabolism Baculoviridae/genetics Base Sequence Binding Sites/genetics Cell Line Cell Transformation, Neoplastic DNA Primers/genetics Mice Mutagenesis, Site-Directed Phosphorylation Polyomavirus/genetics,immunology,metabolism Polyomavirus Infections/etiology Protein Binding Protein-Tyrosine Kinases/genetics,metabolism Proteins/metabolism Serine/chemistry Spodoptera Transfection Tumor Virus Infections/etiology Tyrosine 3-Monooxygenase
Chemicals
14-3-3 Proteins Antigens, Polyomavirus Transforming DNA Primers Proteins Serine Tyrosine 3-Monooxygenase Protein-Tyrosine Kinases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Culleré X
Department of Biochemistry, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
Rose P
Thathamangalam U
Chatterjee A
Mullane K P
Pallas D C
Benjamin T L
Roberts T M
Schaffhausen B S
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.72.1.558-563.1998
Published
1998-01-00
Pages
558-63
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC109408
Subset
IM
Grants
NCI NIH HHS · P01-CA50661 · United States
NCI NIH HHS · R01 CA057327 · United States
NCI NIH HHS · P0-CA50661 · United States
NCI NIH HHS · R37-CA34722 · United States
NCI NIH HHS · R01 CA034722 · United States
NCI NIH HHS · P01 CA050661 · United States
External Links
PubMed source DOI: 10.1128/JVI.72.1.558-563.1998 PMC Full Text View journal details
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