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PMID: 7935389 Published · ppublish English Journal Article

Mechanism of inhibition of Raf-1 by protein kinase A.

Molecular and cellular biology ·Vol. 14 ·No. 10 ·1994-10-00 ·Pages 6696-703

Häfner S, Adler HS, Mischak H, Janosch P, Heidecker G, Wolfman A, Pippig S, Lohse M, Ueffing M, Kolch W

Abstract

The cytoplasmic Raf-1 kinase is essential for mitogenic signalling by growth factors, which couple to tyrosine kinases, and by tumor-promoting phorbol esters such as 12-O-tetradecanoylphorbol-13-acetate, which activate protein kinase C (PKC). Signalling by the Raf-1 kinase can be blocked by activation of the cyclic AMP (cAMP)-dependent protein kinase A (PKA). The molecular mechanism of this inhibition is not precisely known but has been suggested to involve attenuation of Raf-1 binding to Ras. Using purified proteins, we show that in addition to weakening the interaction of Raf-1 with Ras, PKA can inhibit Raf-1 function directly via phosphorylation of the Raf-1 kinase domain. Phosphorylation by PKA interferes with the activation of Raf-1 by either PKC alpha or the tyrosine kinase Lck and even can downregulate the kinase activity of Raf-1 previously activated by PKC alpha or amino-terminal truncation. This type of inhibition can be dissociated from the ability of Raf-1 to associate with Ras, since (i) the isolated Raf-1 kinase domain, which lacks the Ras binding domain, is still susceptible to inhibition by PKA, (ii) phosphorylation of Raf-1 by PKC alpha alleviates the PKA-induced reduction of Ras binding but does not prevent the downregulation of Raf-1 kinase activity by PKA and (iii) cAMP agonists antagonize transformation by v-Raf, which is Ras independent.

MeSH Terms
3T3 Cells Animals Cell Transformation, Neoplastic Cyclic AMP-Dependent Protein Kinases/metabolism Enzyme Activation Isoenzymes/metabolism Lymphocyte Specific Protein Tyrosine Kinase p56(lck) MAP Kinase Kinase Kinase 1 Mice Oncogene Proteins v-raf Phosphorylation Protein Binding Protein Kinase C/metabolism Protein Kinase C-alpha Protein Serine-Threonine Kinases/metabolism Protein-Tyrosine Kinases/metabolism Proto-Oncogene Proteins/metabolism Proto-Oncogene Proteins c-raf Proto-Oncogene Proteins p21(ras)/metabolism Retroviridae Proteins, Oncogenic/metabolism Substrate Specificity
Chemicals
Isoenzymes Proto-Oncogene Proteins Retroviridae Proteins, Oncogenic Protein-Tyrosine Kinases Lymphocyte Specific Protein Tyrosine Kinase p56(lck) Oncogene Proteins v-raf Protein Serine-Threonine Kinases Proto-Oncogene Proteins c-raf Cyclic AMP-Dependent Protein Kinases Prkca protein, mouse Protein Kinase C Protein Kinase C-alpha MAP Kinase Kinase Kinase 1 Map3k1 protein, mouse Proto-Oncogene Proteins p21(ras)
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Häfner S
Institut für Klinische Molekularbiologie und Tumorgenetik, Munich.
Adler H S
Mischak H
Janosch P
Heidecker G
Wolfman A
Pippig S
Lohse M
Ueffing M
Kolch W
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47 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.10.6696-6703.1994
Published
1994-10-00
Pages
6696-703
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359200
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.14.10.6696-6703.1994 PMC Full Text View journal details
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