Myosin Vb interacts with Rab8a on a tubular network containing EHD1 and EHD3.
Cells use multiple pathways to internalize and recycle cell surface components. Although Rab11a and Myosin Vb are involved in the recycling of proteins internalized by clathrin-mediated endocytosis, Rab8a has been implicated in nonclathrin-dependent endocytosis and recycling. By yeast two-hybrid assays, we have now demonstrated that Myosin Vb can interact with Rab8a, but not Rab8b. We have confirmed the interaction of Myosin Vb with Rab11a and Rab8a in vivo by using fluorescent resonant energy transfer techniques. Rab8a and Myosin Vb colocalize to a tubular network containing EHD1 and EHD3, which does not contain Rab11a. Myosin Vb tail can cause the accumulation of both Rab11a and Rab8a in collapsed membrane cisternae, whereas dominant-negative Rab11-FIP2(129-512) selectively accumulates Rab11a but not Rab8a. Additionally, dynamic live cell imaging demonstrates distinct pathways for Rab11a and Rab8a vesicle trafficking. These findings indicate that Rab8a and Rab11a define different recycling pathways that both use Myosin Vb.
Roland
Joseph T
JT
Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN 37232-2733, USA.
Kenworthy
Anne K
AK
Peranen
Johan
J
Caplan
Steve
S
Goldenring
James R
JR
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002645:Chickens; D031541:Fluorescence Resonance Energy Transfer; D006367:HeLa Cells; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D024701:Myosin Type V / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D011817:Rabbits; D022161:Transport Vesicles / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D033921:Vesicular Transport Proteins / Q000378:metabolism*; D020691:rab GTP-Binding Proteins / Q000378:metabolism*; D000099193:rab11 GTP-Binding Proteins
D002352:Carrier Proteins; D015395:Histocompatibility Antigens Class I; D024701:Myosin Type V; D033921:Vesicular Transport Proteins; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins; C120700:EHD1 protein, human; C406446:EHD2 protein, human; C082377:RAB8A protein, human