GAP-independent termination of photoreceptor light response by excess gamma subunit of the cGMP-phosphodiesterase.
We have generated a mouse with rod photoreceptors overexpressing the gamma inhibitory subunit (PDE6gamma) of the photoreceptor G-protein effector cGMP phosphodiesterase (PDE6). PDE6gamma overexpression decreases the rate of rise of the rod response at dim intensities, indicating a reduction in the gain of transduction that may be the result of cytoplasmic PDE6gamma binding to activated transducin alpha GTP (Talpha-GTP) before the Talpha-GTP binds to endogenous PDE6gamma. Excess PDE6gamma also produces a marked acceleration in the falling phase of the light response and more rapid recovery of sensitivity and circulating current after prolonged light exposure. These effects are not mediated by accelerating GTP hydrolysis through the GAP (GTPase activating protein) complex, because the decay of the light response is also accelerated in rods that overexpress PDE6gamma but lack RGS9. Our results show that the PDE6gamma binding sites of PDE6 alpha and beta are accessible to excess (presumably cytoplasmic) PDE6gamma in the light, once endogenous PDE6gamma has been displaced from its binding site by Talpha-GTP. They also suggest that in the presence of Talpha-GTP, the PDE6gamma remains attached to the rest of the PDE6 molecule, but after conversion of Talpha-GTP to Talpha-GDP, the PDE6gamma may dissociate from the PDE6 and exchange with a cytoplasmic pool. This pool may exist even in wild-type rods and may explain the decay of rod photoresponses in the presence of nonhydrolyzable analogs of GTP.
Tsang
Steven H
SH
Department of Pathology, Center for Neurobiology and Behavior, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
Woodruff
Michael L
ML
Chen
Ching-Kang
CK
Yamashita
Clyde Y
CY
Cilluffo
Marianne C
MC
Rao
Anjali L
AL
Farber
Debora B
DB
Fain
Gordon L
GL
D000818:Animals; D002478:Cells, Cultured; D054707:Cyclic Nucleotide Phosphodiesterases, Type 6; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D008027:Light; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009865:Oocytes / Q000502:physiology* / Q000528:radiation effects; D010727:Phosphoric Diester Hydrolases / Q000235:genetics / Q000378:metabolism*; D021122:Protein Subunits; D017948:Retinal Rod Photoreceptor Cells / Q000502:physiology* / Q000528:radiation effects
D020690:GTPase-Activating Proteins; D021122:Protein Subunits; D010727:Phosphoric Diester Hydrolases; D054707:Cyclic Nucleotide Phosphodiesterases, Type 6; C517438:Pde6b protein, mouse; C517444:Pde6g protein, mouse