Abstract
Subcellular localization directed by specific A kinase anchoring proteins (AKAPs) is a mechanism for compartmentalization of cAMP-dependent protein kinase (PKA). Using a two-hybrid screen, a novel AKAP was isolated. Because it interacts with both the type I and type II regulatory subunits, it was defined as a dual specific AKAP or D-AKAP1. Here we report the cloning and characterization of another novel cDNA isolated from that screen. This new member of the D-AKAP family, D-AKAP2, also binds both types of regulatory subunits. A message of 5 kb pairs was detected for D-AKAP2 in all embryonic stages and in all adult tissues tested. In brain, skeletal muscle, kidney, and testis, a 10-kb mRNA was identified. In testis, several small mRNAs were observed. Therefore, D-AKAP2 represents a novel family of proteins. cDNA cloning from a mouse testis library identified the full length D-AKAP2. It is composed of 372 amino acids which includes the R binding fragment, residues 333-372, at its C-terminus. Based on coprecipitation assays, the R binding domain interacts with the N-terminal dimerization domain of RIalpha and RIIalpha. A putative RGS domain was identified near the N-terminal region of D-AKAP2. The presence of this domain raises the intriguing possibility that D-AKAP2 may interact with a Galpha protein thus providing a link between the signaling machinery at the plasma membrane and the downstream kinase.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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A Kinase Anchor Proteins
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Adaptor Proteins, Signal Transducing*
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Amino Acid Sequence
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Animals
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Base Sequence
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Binding Sites
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Carrier Proteins / biosynthesis
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Carrier Proteins / chemistry*
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Carrier Proteins / metabolism*
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Cloning, Molecular
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Conserved Sequence
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Cyclic AMP-Dependent Protein Kinase RIIalpha Subunit
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Cyclic AMP-Dependent Protein Kinase RIalpha Subunit
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Cyclic AMP-Dependent Protein Kinase Type II
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Cyclic AMP-Dependent Protein Kinases / biosynthesis
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Cyclic AMP-Dependent Protein Kinases / chemistry*
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Cyclic AMP-Dependent Protein Kinases / metabolism*
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DNA, Complementary
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Embryo, Mammalian
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GTPase-Activating Proteins
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Gene Library
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Macromolecular Substances
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Male
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Mice
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Molecular Sequence Data
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Organ Specificity
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Proteins*
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RNA, Messenger / biosynthesis
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Sequence Alignment
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Sequence Homology, Amino Acid
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Substrate Specificity
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Testis / enzymology
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Transcription, Genetic
Substances
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A Kinase Anchor Proteins
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Adaptor Proteins, Signal Transducing
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Akap10 protein, mouse
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Carrier Proteins
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Cyclic AMP-Dependent Protein Kinase RIIalpha Subunit
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Cyclic AMP-Dependent Protein Kinase RIalpha Subunit
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DNA, Complementary
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GTPase-Activating Proteins
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Macromolecular Substances
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Prkar1a protein, mouse
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Prkar2a protein, mouse
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Proteins
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RNA, Messenger
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Recombinant Proteins
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Cyclic AMP-Dependent Protein Kinase Type II
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Cyclic AMP-Dependent Protein Kinases