Receptor-mediated Ca2+ and PKC signaling triggers the loss of cortical PKA compartmentalization through the redistribution of gravin

Micah B. Schott, Bryon Grove

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A-Kinase Anchoring Proteins (AKAPs) direct the flow of cellular information by positioning multiprotein signaling complexes into proximity with effector proteins. However, certain AKAPs are not stationary but can undergo spatiotemporal redistribution in response to stimuli. Gravin, a 300kD AKAP that intersects with a diverse signaling array, is localized to the plasma membrane but has been shown to translocate to the cytosol following the elevation of intracellular calcium ([Ca2+]i). Despite the potential for gravin redistribution to impact multiple signaling pathways, the dynamics of this event remain poorly understood. In this study, quantitative microscopy of cells expressing gravin-EGFP revealed that Ca2+ elevation caused the complete translocation of gravin from the cell cortex to the cytosol in as little as 60s of treatment with ionomycin or thapsigargin. In addition, receptor mediated signaling was also shown to cause gravin redistribution following ATP treatment, and this event required both [Ca2+]i elevation and PKC activation. To understand the mechanism for Ca2+ mediated gravin dynamics, deletion of calmodulin-binding domains revealed that a fourth putative calmodulin binding domain called CB4 (a.a. 670-694) is critical for targeting gravin to the cell cortex despite its location downstream of gravin's membrane-targeting domains, which include an N-terminal myristoylation site and three polybasic domains. Finally, confocal microscopy of cells co-transfected with gravin-EYFP and PKA RII-ECFP revealed that gravin redistribution mediated by ionomycin, thapsigargin, and ATP each triggered the gravin-dependent loss of PKA localized at the cell cortex. Our results support the hypothesis that gravin redistribution regulates cross-talk between PKA-dependent signaling and receptor-mediated events involving Ca2+ and PKC.

Original languageEnglish (US)
Pages (from-to)2125-2135
Number of pages11
JournalCellular Signalling
Volume25
Issue number11
DOIs
StatePublished - Nov 1 2013

Fingerprint

Protein Kinases
Ionomycin
Thapsigargin
Calmodulin
Cytosol
Adenosine Triphosphate
Multiprotein Complexes
Confocal Microscopy
Microscopy
Cell Membrane
Calcium
Membranes
Proteins

Keywords

  • AKAP
  • AKAP12
  • Calcium
  • Gravin
  • PKA
  • Purinergic receptor

ASJC Scopus subject areas

  • Cell Biology

Cite this

Receptor-mediated Ca2+ and PKC signaling triggers the loss of cortical PKA compartmentalization through the redistribution of gravin. / Schott, Micah B.; Grove, Bryon.

In: Cellular Signalling, Vol. 25, No. 11, 01.11.2013, p. 2125-2135.

Research output: Contribution to journalArticle

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