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PMID: 7991605 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Oligodeoxynucleotides antisense to mRNA encoding protein kinase A, protein kinase C, and beta-adrenergic receptor kinase reveal distinctive cell-type-specific roles in agonist-induced desensitization.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 91 ·No. 25 ·1994-12-06 ·Pages 12193-7

Shih M, Malbon CC

Abstract

The roles of three protein kinases, cyclic AMP-dependent protein kinase (protein kinase A), protein kinase C, and beta-adrenergic receptor kinase (beta ARK), implicated in agonist-induced desensitization of guanine nucleotide-binding protein (G-protein)-coupled receptors were explored in four different cell lines after 48 hr of incubation with oligodeoxynucleotides antisense to the mRNA encoding each kinase. Desensitization of beta 2-adrenergic receptors was analyzed in cell types in which the activities of the endogenous complement of protein kinases A and C and beta ARK were distinctly different. Protein kinase A was necessary for desensitization of rat osteosarcoma cells (ROS 17/2.8), whereas the contribution of beta ARK to desensitization was insignificant. In Chinese hamster ovary cells that stably express beta 2-adrenergic receptors and in smooth muscle cells (DDT1MF-2), oligodeoxynucleotides antisense to beta ARK mRNA nearly abolished desensitization, whereas oligodeoxynucleotides antisense to protein kinase A mRNA attenuated desensitization to a lesser extent. In human epidermoid carcinoma cells (A-431), oligodeoxynucleotides antisense to either protein kinase A mRNA or beta ARK mRNA attenuated agonist-induced desensitization, providing a third scenario in which two kinases constitute the basis for agonist-induced desensitization. In sharp contrast, oligodeoxynucleotides antisense to protein kinase C mRNA were found to enhance rather than attenuate desensitization in DDT1MF-2 and A-431 cell lines, demonstrating counterregulation between prominent protein kinases in desensitization. Using antisense oligodeoxynucleotides to "knock out" target protein kinases in vivo, we reveal distinctive cell-type-specific roles of protein kinase A, protein kinase C, and beta ARK in agonist-induced desensitization.

MeSH Terms
Animals Base Sequence CHO Cells Cell Line Cricetinae Cyclic AMP/metabolism Cyclic AMP-Dependent Protein Kinases/biosynthesis Isoproterenol/pharmacology Male Molecular Sequence Data Muscle, Smooth Oligonucleotides, Antisense/pharmacology Osteosarcoma Protein Kinase C/biosynthesis RNA, Messenger/metabolism Rats Receptors, Adrenergic, beta-2/drug effects,physiology Transfection Tumor Cells, Cultured Vas Deferens beta-Adrenergic Receptor Kinases
Chemicals
Oligonucleotides, Antisense RNA, Messenger Receptors, Adrenergic, beta-2 Cyclic AMP Cyclic AMP-Dependent Protein Kinases Protein Kinase C beta-Adrenergic Receptor Kinases Isoproterenol
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Shih M
Department of Molecular Pharmacology, School of Medicine, State University of New York, Stony Brook 11794-8651.
Malbon C C
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.91.25.12193
Published
1994-12-06
Pages
12193-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC45403
Subset
IM
Grants
NIDDK NIH HHS · DK25410 · United States
External Links
PubMed source DOI: 10.1073/pnas.91.25.12193 PMC Full Text View journal details
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