biostudies-literatureRoland JTNEI NIH HHSNICHD NIH HHSNIDDK NIH HHSNCI NIH HHSNIGMS NIH HHS2828-37https://www.ebi.ac.uk/biostudies/studies/S-EPMC1949367biostudies-literatureUnknown18(8)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.Molecular biology of the cellMyosin Vb interacts with Rab8a on a tubular network containing EHD1 and EHD3.PMC1949367CA-68485P30 DK058404HD-15052R01 GM073846-02R01 DK070856R01 DK048370R01 GM073846-03P30 EY008126R01GM-073846EY-08126P30 DK020593R01 GM073846-01A1P30 HD015052R01 GM073846DK-59637U24 DK059637P60 DK020593F32DK-072789DK-20593R01DK-48370R01GM-074877P30 CA068485R01DK-070856F32 DK072789DK-58404Roland JTCaplan SGoldenring JRKenworthy AKPeranen JfalseMyosin 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.2007-01-01T00:00:00Z2007 Aug2022-02-09T17:00:15.056Z2019-03-27T02:21:19ZS-EPMC19493671750764710.1091/mbc.e07-02-0169