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PMID: 8816492 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Direct interaction between protein kinase C theta (PKC theta) and 14-3-3 tau in T cells: 14-3-3 overexpression results in inhibition of PKC theta translocation and function.

Molecular and cellular biology ·Vol. 16 ·No. 10 ·1996-10-00 ·Pages 5782-91

Meller N, Liu YC, Collins TL, Bonnefoy-Bérard N, Baier G, Isakov N, Altman A

Abstract

Recent studies have documented direct interactions between 14-3-3 proteins and several oncogene and proto-oncogene products involved in signal transduction pathways. Studies on the effects of 14-3-3 proteins on protein kinase C (PKC) activity in vitro have reported conflicting results, and previous attempts to demonstrate a direct association between PKC and 14-3-3 were unsuccessful. Here, we examined potential physical and functional interactions between PKC theta, a Ca(2+)-independent PKC enzyme which is expressed selectively in T lymphocytes, and the 14-3-3 tau isoform in vitro and in intact T cells. PKC theta and 14-3-3 tau coimmunoprecipitated from Jurkat T cells, and recombinant 14-3-3 tau interacted directly with purified PKC theta in vitro. Transient overexpression of 14-3-3 tau suppressed stimulation of the interleukin 2 (IL-2) promoter mediated by cotransfected wild-type or constitutively active PKC theta, as well as by endogenous PKC in ionomycin- and/or phorbol ester-stimulated cells. This did not represent a general inhibition of activation events, since PKC-independent (but Ca(2+)-dependent) activation of an IL-4 promoter element was not inhibited by 14-3-3 tau under similar conditions. Overexpression of wild-type 14-3-3 tau also inhibited phorbol ester-induced PKC theta translocation from the cytosol to the membrane in Jurkat cells, while a membrane-targeted form of 14-3-3 tau caused increased localization of PKC theta in the particulate fraction in unstimulated cells. Membrane-targeted 14-3-3 tau was more effective than wild-type 14-3-3 tau in suppressing PKC theta-dependent IL-2 promoter activity, suggesting that 14-3-3 tau inhibits the function of PKC theta not only by preventing its translocation to the membrane but also by associating with it. The interaction between 14-3-3 and PKC theta may represent an important general mechanism for regulating PKC-dependent signals and, more specifically, PKC theta-mediated functions during T-cell activation.

MeSH Terms
14-3-3 Proteins Blotting, Western Chloramphenicol O-Acetyltransferase/biosynthesis Gene Expression Regulation Glutathione Transferase/biosynthesis Humans Interleukin-2/biosynthesis,genetics Ionomycin/pharmacology Isoenzymes/biosynthesis,isolation & purification,metabolism Jurkat Cells Mutagenesis, Site-Directed Point Mutation Promoter Regions, Genetic Protein Biosynthesis Protein Kinase C/biosynthesis,isolation & purification,metabolism Protein Kinase C-theta Proteins/isolation & purification,metabolism Proto-Oncogene Mas Recombinant Proteins/biosynthesis,isolation & purification,metabolism Sequence Tagged Sites Signal Transduction T-Lymphocytes/metabolism Tetradecanoylphorbol Acetate/pharmacology Transfection Tyrosine 3-Monooxygenase
Chemicals
14-3-3 Proteins Interleukin-2 Isoenzymes MAS1 protein, human Proteins Proto-Oncogene Mas Recombinant Proteins Ionomycin Tyrosine 3-Monooxygenase Chloramphenicol O-Acetyltransferase Glutathione Transferase PRKCQ protein, human Protein Kinase C Protein Kinase C-theta Tetradecanoylphorbol Acetate
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Meller N
Division of Cell Biology, La Jolla Institute for Allergy and Immunology, San Diego, California 92121, USA.
Liu Y C
Collins T L
Bonnefoy-Bérard N
Baier G
Isakov N
Altman A
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.10.5782
Published
1996-10-00
Pages
5782-91
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231579
Subset
IM
Grants
NCI NIH HHS · CA35299 · United States
External Links
PubMed source DOI: 10.1128/MCB.16.10.5782 PMC Full Text View journal details
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