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J Cell Sci. 2008 May 1;121(Pt 9):1363-72. doi: 10.1242/jcs.020081. Epub 2008 Apr 8.

Caveolin-1 alters Ca(2+) signal duration through specific interaction with the G alpha q family of G proteins.

Author information

1
Department of Physiology and Biophysics, BST6-145, Stony Brook University, Stony Brook, NY 11794-8661, USA.

Abstract

Caveolae are membrane domains having caveolin-1 (Cav1) as their main structural component. Here, we determined whether Cav1 affects Ca(2+) signaling through the Galpha(q)-phospholipase-Cbeta (PLCbeta) pathway using Fischer rat thyroid cells that lack Cav1 (FRTcav(-)) and a sister line that forms caveolae-like domains due to stable transfection with Cav1 (FRTcav(+)). In the resting state, we found that eCFP-Gbetagamma and Galpha(q)-eYFP are similarly associated in both cell lines by Forster resonance energy transfer (FRET). Upon stimulation, the amount of FRET between Galpha(q)-eYFP and eCFP-Gbetagamma remains high in FRTcav(-) cells, but decreases almost completely in FRTcav(+) cells, suggesting that Cav1 is increasing the separation between Galpha(q)-Gbetagamma subunits. In FRTcav(-) cells overexpressing PLCbeta, a rapid recovery of Ca(2+) is observed after stimulation. However, FRTcav(+) cells show a sustained level of elevated Ca(2+). FRET and colocalization show specific interactions between Galpha(q) and Cav1 that increase upon stimulation. Fluorescence correlation spectroscopy studies show that the mobility of Galpha(q)-eGFP is unaffected by activation in either cell type. The mobility of eGFP-Gbetagamma remains slow in FRTcav(-) cells but increases in FRTcav(+) cells. Together, our data suggest that, upon stimulation, Galpha(q)(GTP) switches from having strong interactions with Gbetagamma to Cav1, thereby releasing Gbetagamma. This prolongs the recombination time for the heterotrimer, thus causing a sustained Ca(2+) signal.

PMID:
18397999
PMCID:
PMC2639659
DOI:
10.1242/jcs.020081
[Indexed for MEDLINE]
Free PMC Article

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