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First published online May 21, 2004; 10.1105/tpc.020321

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The Plant Cell 16:1616-1632 (2004)
© 2004 American Society of Plant Biologists

The Arabidopsis Putative G Protein–Coupled Receptor GCR1 Interacts with the G Protein {alpha} Subunit GPA1 and Regulates Abscisic Acid Signaling

Sona Pandey and Sarah M. Assmann1

Biology Department, Penn State University, University Park, Pennsylvania, 16802

1 To whom correspondence should be addressed. E-mail sma3{at}psu.edu; fax 814-865-9131.

Heterotrimeric G proteins composed of {alpha}, ß, and {gamma} subunits link ligand perception by G protein–coupled receptors (GPCRs) with downstream effectors, providing a ubiquitous signaling mechanism in eukaryotes. The Arabidopsis thaliana genome encodes single prototypical G{alpha} (GPA1) and Gß (AGB1) subunits, and two probable G{gamma} subunits (AGG1 and AGG2). One Arabidopsis gene, GCR1, encodes a protein with significant sequence similarity to nonplant GPCRs and a predicted 7-transmembrane domain structure characteristic of GPCRs. However, whether GCR1 actually interacts with GPA1 was unknown. We demonstrate by in vitro pull-down assays, by yeast split-ubiquitin assays, and by coimmunoprecipitation from plant tissue that GCR1 and GPA1 are indeed physically coupled. GCR1–GPA1 interaction depends on intracellular domains of GCR1. gcr1 T-DNA insertional mutants exhibit hypersensitivity to abscisic acid (ABA) in assays of root growth, gene regulation, and stomatal response. gcr1 guard cells are also hypersensitive to the lipid metabolite, sphingosine-1-phosphate (S1P), which is a transducer of the ABA signal upstream of GPA1. Because gpa1 mutants exhibit insensitivity in aspects of guard cell ABA and S1P responses, whereas gcr1 mutants exhibit hypersensitivity, GCR1 may act as a negative regulator of GPA1-mediated ABA responses in guard cells.


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