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PMID: 9710607 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

14-3-3 proteins are required for maintenance of Raf-1 phosphorylation and kinase activity.

Molecular and cellular biology ·Vol. 18 ·No. 9 ·1998-09-00 ·Pages 5229-38

Thorson JA, Yu LW, Hsu AL, Shih NY, Graves PR, Tanner JW, Allen PM, Piwnica-Worms H, Shaw AS

Abstract

By binding to serine-phosphorylated proteins, 14-3-3 proteins function as effectors of serine phosphorylation. The exact mechanism of their action is, however, still largely unknown. Here we demonstrate a requirement for 14-3-3 for Raf-1 kinase activity and phosphorylation. Expression of dominant negative forms of 14-3-3 resulted in the loss of a critical Raf-1 phosphorylation, while overexpression of 14-3-3 resulted in enhanced phosphorylation of this site. 14-3-3 levels, therefore, regulate the stoichiometry of Raf-1 phosphorylation and its potential activity in the cell. Phosphorylation of Raf-1, however, was insufficient by itself for kinase activity. Removal of 14-3-3 from phosphorylated Raf abrogated kinase activity, whereas addition of 14-3-3 restored it. This supports a paradigm in which the effects of phosphorylation on serine as well as tyrosine residues are mediated by inducible protein-protein interactions.

MeSH Terms
14-3-3 Proteins Amino Acid Sequence Animals Binding Sites Cell Line Chloramphenicol O-Acetyltransferase/biosynthesis Cloning, Molecular Glutathione Transferase Humans Mice Models, Molecular Mutagenesis, Site-Directed Phosphorylation Phosphoserine Phosphotyrosine Polymerase Chain Reaction Protein Biosynthesis Protein Structure, Secondary Proteins/chemistry,metabolism Proto-Oncogene Proteins c-raf/chemistry,metabolism Recombinant Fusion Proteins/biosynthesis,metabolism Spectrum Analysis Transfection Tyrosine 3-Monooxygenase
Chemicals
14-3-3 Proteins Proteins Recombinant Fusion Proteins Phosphoserine Phosphotyrosine Tyrosine 3-Monooxygenase Chloramphenicol O-Acetyltransferase Glutathione Transferase Proto-Oncogene Proteins c-raf
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Thorson J A
Center for Immunology and Department of Pathology, Washington University School of Medicine, St. Louis, Missouri, USA.
Yu L W
Hsu A L
Shih N Y
Graves P R
Tanner J W
Allen P M
Piwnica-Worms H
Shaw A S
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.18.9.5229
Published
1998-09-00
Pages
5229-38
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC109108
Subset
IM
Grants
NIAID NIH HHS · AI54094 · United States
NIGMS NIH HHS · GM47017 · United States
NIGMS NIH HHS · GM18428 · United States
External Links
PubMed source DOI: 10.1128/MCB.18.9.5229 PMC Full Text View journal details
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