title: Interaction of transducin with uncoordinated 119 protein (UNC119): implications for the model of transducin trafficking in rod photoreceptors.
abstract: The key visual G protein, transducin undergoes bi-directional translocations between the outer segment (OS) and inner compartments of rod photoreceptors in a light-dependent manner thereby contributing to adaptation and neuroprotection of rods. A mammalian uncoordinated 119 protein (UNC119), also known as Retina Gene 4 protein (RG4), has been recently implicated in transducin transport to the OS in the dark through its interaction with the N-acylated GTP-bound transducin-α subunit (Gα(t1)). Here, we demonstrate that the interaction of human UNC119 (HRG4) with transducin is dependent on the N-acylation, but does not require the GTP-bound form of Gα(t1). The lipid specificity of UNC119 is unique: UNC119 bound the myristoylated N terminus of Gα(t1) with much higher affinity than a prenylated substrate, whereas the homologous prenyl-binding protein PrBP/δ did not interact with the myristoylated peptide. UNC119 was capable of interacting with Gα(t1)GDP as well as with heterotrimeric transducin (G(t)). This interaction of UNC119 with G(t) led to displacement of Gβ(1)γ(1) from the heterotrimer. Furthermore, UNC119 facilitated solubilization of G(t) from dark-adapted rod OS membranes. Consistent with these observations, UNC119 inhibited rhodopsin-dependent activation of G(t), but had no effect on the GTP-hydrolysis by Gα(t1). A model for the role of UNC119 in the IS→OS translocation of G(t) is proposed based on the UNC119 ability to dissociate G(t) subunits from each other and the membrane. We also found that UNC119 inhibited activation of G(o) by D2 dopamine receptor in cultured cells. Thus, UNC119 may play conserved inhibitory role in regulation of GPCR-G protein signaling in non-visual tissues.
authors.affiliation: Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, Iowa 52242.
authors.forename: Kota N
authors.initials: KN
authors.lastname: Gopalakrishna
authors.affiliation: Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, Iowa 52242.
authors.forename: Krishnarao
authors.initials: K
authors.lastname: Doddapuneni
authors.affiliation: Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, Iowa 52242.
authors.forename: Kimberly K
authors.initials: KK
authors.lastname: Boyd
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, Iowa 52242 and.
authors.forename: Ikuo
authors.initials: I
authors.lastname: Masuho
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, Iowa 52242 and.
authors.forename: Kirill A
authors.initials: KA
authors.lastname: Martemyanov
authors.affiliation: Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, Iowa 52242; Department of Neuroscience, The Scripps Research Institute, Jupiter, Florida 33458. Electronic address: nikolai-artemyev@uiowa.edu.
authors.forename: Nikolai O
authors.initials: NO
authors.lastname: Artemyev
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism*; D020962:Heterotrimeric GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D021381:Protein Transport / Q000502:physiology; D011954:Receptors, Dopamine / Q000235:genetics / Q000378:metabolism; D017948:Retinal Rod Photoreceptor Cells / Q000166:cytology / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D015293:Transducin / Q000235:genetics / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D011954:Receptors, Dopamine; C098032:UNC119 protein, human; D020962:Heterotrimeric GTP-Binding Proteins; D015293:Transducin