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Endocrinology Vol. 139, No. 6 2784-2795
Copyright © 1998 by The Endocrine Society

ARTICLES

Rat G Protein-Coupled Receptor Kinase GRK4: Identification, Functional Expression, and Differential Tissue Distribution of Two Splice Variants1

Bérangère Virlon2, Dmitri Firsov2,3, Lydie Cheval, Eric Reiter, Carine Troispoux4, Florian Guillou and Jean-Marc Elalouf

Département de Biologie Cellulaire et Moléculaire (B.V., D.F., L.C., J.-M.E.), Service de Biologie Cellulaire, CEA Saclay, 91191 Gif-sur-Yvette Cedex, France; Institut National de la Recherche Agronomique (INRA) (E.R., C.T., F.G.), Station de Physiologie de la Reproduction des Mammifères Domestiques, URA CNRS 1291, 37380 Nouzilly, France

Address all correspondence and requests for reprints to: J. M. Elalouf, Département de Biologie Cellulaire et Moléculaire, Service de Biologie Cellulaire, CEA SACLAY, 91191 Gif-sur-Yvette Cedex, France. E-mail: elalouf{at}dsvidf.cea.fr

G protein-coupled receptor kinases (GRKs) specifically phosphorylate the agonist-occupied form of G protein-coupled receptors, leading to the homologous mode of desensitization. We report here on the cloning of complementary DNAs that encode two rat GRK4 variants. Rat GRK4A (575 amino acids) displays 76% identity with the long human GRK4 splice variant. Rat GRK4B (545 amino acids) delineates a new variant that is identical to GRK4A except for a 31-amino acid deletion in the N-terminal domain, corresponding to exon VI in the human GRK4 gene. GRKs4A and B are likely produced by alternative splicing from a single gene, the partial characterization of which revealed a structural organization similar to that of the human GRK4 gene. GRK4A messenger RNA (mRNA) is abundant only in testis. A combination of in situ hybridization and quantitative RT-PCR studies demonstrated that GRK4A mRNA level increases during testicular development and predominates in leptotene to late pachytene primary spermatocytes and round spermatids. GRK4B mRNA is poorly expressed in testis and most rat tissues but is heterogeneously distributed in the kidney, with 20-fold enrichment in the outer medulla. GRKs4A and B are both functional protein kinases, as demonstrated in a rhodopsin phosphorylation assay. The differential tissue distribution of GRKA4 and GRK4B suggests that individual GRK4 variants may serve distinct physiological functions.




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