Rab6 is associated with a compartment that transports rhodopsin from the trans-Golgi to the site of rod outer segment disk formation in frog retinal photoreceptors.
The biogenesis of light sensitive membranes in retinal rod photoreceptors involves polarized sorting and targeting of newly synthesized rhodopsin to a specialized domain, the rod outer segment (ROS). We have isolated and characterized the population of post-Golgi membranes that mediate intracellular transport of rhodopsin. In the present study we have examined the association of small (20-25 kDa) GTP-binding (G) proteins with these membranes. We found that one of the small G proteins, rab6, behaves like an integral membrane protein of the post-Golgi vesicles, although approximately 30% of rab6 is soluble. The distribution of the membrane-associated and the soluble forms is highly polarized. By confocal and EM immunocytochemistry it can be seen that most of rab6 is associated with the photoreceptor trans-Golgi cisternae, trans-Golgi network (TGN) and post-Golgi vesicles. The photoreceptor axon and synaptic terminal are unlabeled, but dendrites of deeper retinal layers are labeled. The distribution of rab6 across sucrose density gradient fractions parallels the distribution of sialyltransferase (a TGN marker) activity. About 9% of membrane-bound rab6 is associated, however, with the rhodopsin-bearing sialyltransferase-free post-Golgi vesicles, which represent a very small fraction (< 1%) of the total retinal membranes. Rab6 is absent from the mature ROS disk membranes but it is present at the sites of new ROS disk formation and in the ROS cytoplasm. This suggests that rab6 becomes soluble upon disk membrane formation. Therefore, rab6 may function not only as a component of the sorting machinery of photoreceptors that delivers rhodopsin to its appropriate subcellular domain but may also participate in some aspects of ROS disk morphogenesis.
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Department of Pathology, University of Texas Health Science Center at San Antonio 78284-7750.
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D000818:Animals; D001692:Biological Transport; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D011898:Ranidae; D017948:Retinal Rod Photoreceptor Cells / Q000378:metabolism*; D012243:Rhodopsin / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism
D012243:Rhodopsin; D019204:GTP-Binding Proteins