Inhibitory cross-talk by cAMP kinase on the calmodulin-dependent protein kinase cascade

Gary A. Wayman, Hiroshi Tokumitsu, Thomas R. Soderling

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

The calmodulin-dependent kinase (CaM-K) cascade, a Ca2+-triggered system involving phosphorylation and activation of CaM-KI and CaM-KIV by CaM kinase kinase (CaM-KK), regulates transcription through direct phosphorylation of transcription factors such as cAMP response element- binding protein. We have shown previously that activated CaM-KIV can activate the mitogen-activated protein kinases (Enslen, H., Tokumitsu, H., Stork, P. J. S., Davis, R. J., and Soderling, T. R. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 10803-10808), and the present paper describes a novel regulatory cross-talk between cAMP kinase (PKA) and CaM-KK. PKA gave rapid phosphorylation in vitro and in cells of recombinant CaM-KK, resulting in 50- 75% inhibition of CaM-KK activity, part of which was due to suppression of CaM-binding by phosphorylation of Ser458 in the CaM-binding domain. However, the Ser458 Ala mutant, or a truncation mutant in which the CaM- binding and autoinhibitory domains were deleted, was still partially suppressed by PKA-mediated phosphorylation. The second inhibitory site was identified as Thr108 by site-specific mutagenesis. Treatments of COS-7, PC12, hippocampal, or Jurkat cells with the PKA activators forskolin or isoproterenol gave 30-90% inhibition of either endogenous or transfected CaM- KK and/or CaM-KIV activities. These results demonstrate that the CaM kinase cascade is negatively regulated in cells by the cAMP/PKA pathway.

Original languageEnglish
Pages (from-to)16073-16076
Number of pages4
JournalJournal of Biological Chemistry
Volume272
Issue number26
DOIs
Publication statusPublished - Jun 27 1997
Externally publishedYes

Fingerprint

Calcium-Calmodulin-Dependent Protein Kinases
Phosphotransferases
Phosphorylation
Calcium-Calmodulin-Dependent Protein Kinase Kinase
Cyclic AMP Response Element-Binding Protein
Jurkat Cells
PC12 Cells
Mutagenesis
Colforsin
Site-Directed Mutagenesis
Mitogen-Activated Protein Kinases
Isoproterenol
Transcription
Calmodulin
Transcription Factors
Chemical activation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Inhibitory cross-talk by cAMP kinase on the calmodulin-dependent protein kinase cascade. / Wayman, Gary A.; Tokumitsu, Hiroshi; Soderling, Thomas R.

In: Journal of Biological Chemistry, Vol. 272, No. 26, 27.06.1997, p. 16073-16076.

Research output: Contribution to journalArticle

@article{62bc2dbe315948abb7919ec88048480c,
title = "Inhibitory cross-talk by cAMP kinase on the calmodulin-dependent protein kinase cascade",
abstract = "The calmodulin-dependent kinase (CaM-K) cascade, a Ca2+-triggered system involving phosphorylation and activation of CaM-KI and CaM-KIV by CaM kinase kinase (CaM-KK), regulates transcription through direct phosphorylation of transcription factors such as cAMP response element- binding protein. We have shown previously that activated CaM-KIV can activate the mitogen-activated protein kinases (Enslen, H., Tokumitsu, H., Stork, P. J. S., Davis, R. J., and Soderling, T. R. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 10803-10808), and the present paper describes a novel regulatory cross-talk between cAMP kinase (PKA) and CaM-KK. PKA gave rapid phosphorylation in vitro and in cells of recombinant CaM-KK, resulting in 50- 75{\%} inhibition of CaM-KK activity, part of which was due to suppression of CaM-binding by phosphorylation of Ser458 in the CaM-binding domain. However, the Ser458 Ala mutant, or a truncation mutant in which the CaM- binding and autoinhibitory domains were deleted, was still partially suppressed by PKA-mediated phosphorylation. The second inhibitory site was identified as Thr108 by site-specific mutagenesis. Treatments of COS-7, PC12, hippocampal, or Jurkat cells with the PKA activators forskolin or isoproterenol gave 30-90{\%} inhibition of either endogenous or transfected CaM- KK and/or CaM-KIV activities. These results demonstrate that the CaM kinase cascade is negatively regulated in cells by the cAMP/PKA pathway.",
author = "Wayman, {Gary A.} and Hiroshi Tokumitsu and Soderling, {Thomas R.}",
year = "1997",
month = "6",
day = "27",
doi = "10.1074/jbc.272.26.16073",
language = "English",
volume = "272",
pages = "16073--16076",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "26",

}

TY - JOUR

T1 - Inhibitory cross-talk by cAMP kinase on the calmodulin-dependent protein kinase cascade

AU - Wayman, Gary A.

AU - Tokumitsu, Hiroshi

AU - Soderling, Thomas R.

PY - 1997/6/27

Y1 - 1997/6/27

N2 - The calmodulin-dependent kinase (CaM-K) cascade, a Ca2+-triggered system involving phosphorylation and activation of CaM-KI and CaM-KIV by CaM kinase kinase (CaM-KK), regulates transcription through direct phosphorylation of transcription factors such as cAMP response element- binding protein. We have shown previously that activated CaM-KIV can activate the mitogen-activated protein kinases (Enslen, H., Tokumitsu, H., Stork, P. J. S., Davis, R. J., and Soderling, T. R. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 10803-10808), and the present paper describes a novel regulatory cross-talk between cAMP kinase (PKA) and CaM-KK. PKA gave rapid phosphorylation in vitro and in cells of recombinant CaM-KK, resulting in 50- 75% inhibition of CaM-KK activity, part of which was due to suppression of CaM-binding by phosphorylation of Ser458 in the CaM-binding domain. However, the Ser458 Ala mutant, or a truncation mutant in which the CaM- binding and autoinhibitory domains were deleted, was still partially suppressed by PKA-mediated phosphorylation. The second inhibitory site was identified as Thr108 by site-specific mutagenesis. Treatments of COS-7, PC12, hippocampal, or Jurkat cells with the PKA activators forskolin or isoproterenol gave 30-90% inhibition of either endogenous or transfected CaM- KK and/or CaM-KIV activities. These results demonstrate that the CaM kinase cascade is negatively regulated in cells by the cAMP/PKA pathway.

AB - The calmodulin-dependent kinase (CaM-K) cascade, a Ca2+-triggered system involving phosphorylation and activation of CaM-KI and CaM-KIV by CaM kinase kinase (CaM-KK), regulates transcription through direct phosphorylation of transcription factors such as cAMP response element- binding protein. We have shown previously that activated CaM-KIV can activate the mitogen-activated protein kinases (Enslen, H., Tokumitsu, H., Stork, P. J. S., Davis, R. J., and Soderling, T. R. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 10803-10808), and the present paper describes a novel regulatory cross-talk between cAMP kinase (PKA) and CaM-KK. PKA gave rapid phosphorylation in vitro and in cells of recombinant CaM-KK, resulting in 50- 75% inhibition of CaM-KK activity, part of which was due to suppression of CaM-binding by phosphorylation of Ser458 in the CaM-binding domain. However, the Ser458 Ala mutant, or a truncation mutant in which the CaM- binding and autoinhibitory domains were deleted, was still partially suppressed by PKA-mediated phosphorylation. The second inhibitory site was identified as Thr108 by site-specific mutagenesis. Treatments of COS-7, PC12, hippocampal, or Jurkat cells with the PKA activators forskolin or isoproterenol gave 30-90% inhibition of either endogenous or transfected CaM- KK and/or CaM-KIV activities. These results demonstrate that the CaM kinase cascade is negatively regulated in cells by the cAMP/PKA pathway.

UR - http://www.scopus.com/inward/record.url?scp=0030910235&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030910235&partnerID=8YFLogxK

U2 - 10.1074/jbc.272.26.16073

DO - 10.1074/jbc.272.26.16073

M3 - Article

C2 - 9195898

AN - SCOPUS:0030910235

VL - 272

SP - 16073

EP - 16076

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 26

ER -