An extranuclear locus of cAMP-dependent protein kinase action is necessary and sufficient for promotion of spiral ganglion neuronal survival by cAMP

Jinwoong Bok; Xiang-Ming Zha; Yang-Sun Cho; Steven H Green

doi:10.1523/jneurosci.23-03-00777.2003

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An extranuclear locus of cAMP-dependent protein kinase action is necessary and sufficient for promotion of spiral ganglion neuronal survival by cAMP

Journal article

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Peer reviewed

An extranuclear locus of cAMP-dependent protein kinase action is necessary and sufficient for promotion of spiral ganglion neuronal survival by cAMP

Jinwoong Bok, Xiang-Ming Zha, Yang-Sun Cho and Steven H Green

The Journal of neuroscience, Vol.23(3), pp.777-787

02/01/2003

DOI: 10.1523/jneurosci.23-03-00777.2003

PMCID: PMC6741931

PMID: 12574406

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Abstract

We showed previously that cAMP is a survival-promoting stimulus for cultured postnatal rat spiral ganglion neurons (SGNs) and that depolarization promotes SGN survival in part via recruitment of cAMP signaling. We here investigate the subcellular locus of cAMP prosurvival signaling. Transfection of GPKI, a green fluorescent protein (GFP)-tagged cAMP-dependent protein kinase (PKA) inhibitor, inhibits the ability of the permeant cAMP analog cpt-cAMP [8-(4-chlorophenylthio)-cAMP] to promote survival, indicating that PKA activity is necessary. Transfection of GFP-tagged PKA (GPKA) is sufficient to promote SGN survival, but restriction of GPKA to the nucleus by addition of a nuclear localization signal (GPKAnls) almost completely abrogates its prosurvival effect. In contrast, GPKA targeted to the extranuclear cytoplasm by addition of a nuclear export signal (GPKAnes) promotes SGN survival as effectively as does GPKA. Moreover, GPKI targeted to the nucleus lacks inhibitory effect on SGN survival attributable to cpt-cAMP or depolarization. These data indicate an extranuclear target of PKA for promotion of neuronal survival. Consistent with this, we find that dominant-inhibitory CREB mutants inhibit the prosurvival effect of depolarization but not that of cpt-cAMP. SGN survival is compromised by overexpression of the proapoptotic regulator Bad, previously shown to be phosphorylated in the cytoplasm by PKA. This Bad-induced apoptosis is prevented by cpt-cAMP or by cotransfection of GPKA or of GPKAnes but not of GPKAnls. Thus, cAMP prevents SGN death through a cytoplasmic as opposed to nuclear action, and inactivation of Bad proapoptotic function is a mechanism by which PKA can prevent neuronal death.

Animals

Apoptosis - drug effects

Apoptosis - physiology

bcl-Associated Death Protein

Carrier Proteins - biosynthesis

Carrier Proteins - genetics

Carrier Proteins - pharmacology

Catalytic Domain - genetics

Cell Nucleus - enzymology

Cell Survival - drug effects

Cell Survival - genetics

Cells, Cultured

Cyclic AMP - analogs & derivatives

Cyclic AMP - antagonists & inhibitors

Cyclic AMP - pharmacology

Cyclic AMP Response Element-Binding Protein - metabolism

Cyclic AMP-Dependent Protein Kinases - antagonists & inhibitors

Cyclic AMP-Dependent Protein Kinases - genetics

Cyclic AMP-Dependent Protein Kinases - metabolism

Cytoplasm - enzymology

Enzyme Inhibitors - pharmacology

Green Fluorescent Proteins

Luminescent Proteins - genetics

Neurons - cytology

Neurons - drug effects

Neurons - enzymology

Nuclear Localization Signals - genetics

Nuclear Localization Signals - physiology

Phosphorylation - drug effects

Rats

Recombinant Fusion Proteins - genetics

Recombinant Fusion Proteins - metabolism

Recombinant Fusion Proteins - pharmacology

Signal Transduction - physiology

Spiral Ganglion - cytology

Spiral Ganglion - drug effects

Spiral Ganglion - enzymology

Thionucleotides - antagonists & inhibitors

Thionucleotides - pharmacology

Transfection

Details

Title: Subtitle: An extranuclear locus of cAMP-dependent protein kinase action is necessary and sufficient for promotion of spiral ganglion neuronal survival by cAMP
Creators: Jinwoong Bok - Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242, USA
Xiang-Ming Zha
Yang-Sun Cho
Steven H Green
Resource Type: Journal article
Publication Details: The Journal of neuroscience, Vol.23(3), pp.777-787
DOI: 10.1523/jneurosci.23-03-00777.2003
PMID: 12574406
PMCID: PMC6741931
ISSN: 0270-6474
eISSN: 1529-2401
Grant note: DC02961 / NIDCD NIH HHS R01 DC002961 / NIDCD NIH HHS DK25295 / NIDDK NIH HHS
Language: English
Date published: 02/01/2003
Academic Unit: Biology; Otolaryngology
Record Identifier: 9984217419802771

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