title: Rab GTPase-Myo5B complexes control membrane recycling and epithelial polarization.
abstract: The Rab GTPases are the largest family of proteins regulating membrane traffic. Rab proteins form a nidus for the assembly of multiprotein complexes on distinct vesicle membranes to regulate particular membrane trafficking pathways. Recent investigations have demonstrated that Myosin Vb (Myo5B) is an effector for Rab8a, Rab10, and Rab11a, all of which are implicated in regulating different pathways for recycling of proteins to the plasma membrane. It remains unclear how specific interactions of Myo5B with individual Rab proteins can lead to specificity in the regulation of alternate trafficking pathways. We examined the relative contributions of Rab/Myo5B interactions with specific pathways using Myo5B mutants lacking binding to either Rab11a or Rab8a. Myo5B Q1300L and Y1307C mutations abolished Rab8a association, whereas Myo5B Y1714E and Q1748R mutations uncoupled association with Rab11a. Expression of Myo5B tails containing these mutants demonstrated that Rab11a, but not Rab8a, was required for recycling of transferrin in nonpolarized cells. In contrast, in polarized epithelial cyst cultures, Myo5B was required for apical membrane trafficking and de novo lumen formation, dependent on association with both Rab8a and Rab11a. These data demonstrate that different combinations of Rab GTPase association with Myo5B control distinct membrane trafficking pathways.
authors.affiliation: Department of Surgery, Vanderbilt University School of Medicine, Nashville Veterans Affairs Medical Center Nashville, TN 37232, USA.
authors.forename: Joseph T
authors.initials: JT
authors.lastname: Roland
authors.forename: David M
authors.initials: DM
authors.lastname: Bryant
authors.forename: Anirban
authors.initials: A
authors.lastname: Datta
authors.forename: Aymelt
authors.initials: A
authors.lastname: Itzen
authors.forename: Keith E
authors.initials: KE
authors.lastname: Mostov
authors.forename: James R
authors.initials: JR
authors.lastname: Goldenring
mesh_terms: D000818:Animals; D002460:Cell Line; D016764:Cell Polarity / Q000502:physiology*; D017931:DNA Primers / Q000235:genetics; D004285:Dogs; D004847:Epithelial Cells / Q000502:physiology*; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D007150:Immunohistochemistry; D008566:Membranes / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D046912:Multiprotein Complexes / Q000378:metabolism*; D016296:Mutagenesis; D018995:Myosin Heavy Chains / Q000378:metabolism*; D024701:Myosin Type V / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology; D034622:RNA Interference; D014168:Transferrin / Q000378:metabolism; D022161:Transport Vesicles / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D020691:rab GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D017931:DNA Primers; C505832:MYO5B protein, human; D046912:Multiprotein Complexes; D014168:Transferrin; D024701:Myosin Type V; D018995:Myosin Heavy Chains; D020691:rab GTP-Binding Proteins