Interplay between PIP^sub 3^ and calmodulin regulation of olfactory cyclic nucleotide-gated channels

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 103; no. 42; p. 15635
• Main Authors: Brady, James D, Rich, Elizabeth D, Martens, Jeffrey R, Karpen, Jeffrey W
• Format: Journal Article
• Language: English
• Published: Washington National Academy of Sciences 17.10.2006
• Subjects: Calcium; Neurons; Neurosciences; Proteins
• ISSN: 0027-8424; 1091-6490

Phosphatidylinositol-3,4,5-trisphosphate (PIP^sub 3^) has been proposed to modulate the odorant sensitivity of olfactory sensory neurons by inhibiting activation of cyclic nucleotide-gated (CNG) channels in the cilia. When applied to the intracellular face of excised patches, PIP^sub 3^ has been shown to inhibit activation of heteromeric olfactory CNG channels, composed of CNGA2, CNGA4, and CNGBlb subunits, and homomeric CNGA2 channels. In contrast, we discovered that channels formed by CNGA3 subunits from cone photoreceptors were unaffected by PIP^sub 3^. Using chimeric channels and a deletion mutant, we determined that residues 61-90 within the N terminus of CNGA2 are necessary for PIP^sub 3^ regulation, and a biochemical "pulldown" assay suggests that PIP^sub 3^ directly binds this region. The N terminus of CNGA2 contains a previously identified calcium-calmodulin (Ca2+/CaM)-binding domain (residues 68-81) that mediates Ca2+/CaM inhibition of homomeric CNGA2 channels but is functionally silent in heteromeric channels. We discovered, however, that this region is required for PIP^sub 3^ regulation of both homomeric and heteromeric channels. Furthermore, PIP^sub 3^ occluded the action of Ca2+/CaM on both homomeric and heteromeric channels, in part by blocking Ca2+/CaM binding. Our results establish the importance of the CNGA2 N terminus for PIP^sub 3^ inhibition of olfactory CNG channels and suggest that PIP^sub 3^ inhibits channel activation by disrupting an autoexcitatory interaction between the N and C termini of adjacent subunits. By dramatically suppressing channel currents, PIP^sub 3^ may generate a shift in odorant sensitivity that does not require prior channel activity. [PUBLICATION ABSTRACT]