Unconventional myosin VIIA is a novel A-kinase-anchoring protein.
To gain an insight into the cellular function of the unconventional myosin VIIA, we sought proteins interacting with its tail region, using the yeast two-hybrid system. Here we report on one of the five candidate interactors we identified, namely the type I alpha regulatory subunit (RI alpha) of protein kinase A. The interaction of RI alpha with myosin VIIA tail was demonstrated by coimmunoprecipitation from transfected HEK293 cells. Analysis of deleted constructs in the yeast two-hybrid system showed that the interaction of myosin VIIA with RI alpha involves the dimerization domain of RI alpha. In vitro binding assays identified the C-terminal "4.1, ezrin, radixin, moesin" (FERM)-like domain of myosin VIIA as the interacting domain. In humans and mice, mutations in the myosin VIIA gene underlie hereditary hearing loss, which may or may not be associated with visual deficiency. Immunohistofluorescence revealed that myosin VIIA and RI alpha are coexpressed in the outer hair cells of the cochlea and rod photoreceptor cells of the retina. Our results strongly suggest that myosin VIIA is a novel protein kinase A-anchoring protein that targets protein kinase A to definite subcellular sites of these sensory cells.
Küssel-Andermann
P
P
Unité de Génétique des Déficits Sensoriels, CNRS URA 1968, 25 rue du Dr. Roux, Institut Pasteur, 75724 Paris cedex 15, France.
El-Amraoui
A
A
Safieddine
S
S
Hardelin
J P
JP
Nouaille
S
S
Camonis
J
J
Petit
C
C
D000818:Animals; D001665:Binding Sites; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004398:Dyneins; D004926:Escherichia coli; D006801:Humans; D051379:Mice; D000081412:Myosin VIIa; D009218:Myosins / Q000032:analysis / Q000378:metabolism*; D011485:Protein Binding; D013379:Substrate Specificity
C000631455:MYO7A protein, human; C000631456:Myo7a protein, mouse; D000081412:Myosin VIIa; D017868:Cyclic AMP-Dependent Protein Kinases; D009218:Myosins; D004398:Dyneins