title: Rab11-FIP3 is critical for the structural integrity of the endosomal recycling compartment.
abstract: Rab11-FIP3 is an endosomal recycling compartment (ERC) protein that is implicated in the process of membrane delivery from the ERC to sites of membrane insertion during cell division. Here we report that Rab11-FIP3 is critical for the structural integrity of the ERC during interphase. We demonstrate that knockdown of Rab11-FIP3 and expression of a mutant of Rab11-FIP3 that is Rab11-binding deficient cause loss of all ERC-marker protein staining from the pericentrosomal region of A431 cells. Furthermore, we find that fluorophore-labelled transferrin cannot access the pericentrosomal region of cells in which Rab11-FIP3 function has been perturbed. We find that this Rab11-FIP3 function appears to be specific because expression of the equivalent Rab11-binding deficient mutant of Rab-coupling protein does not perturb ERC morphology. In addition, we find that other organelles such as sorting and late endosomes are unaffected by loss of Rab11-FIP3 function. Finally, we demonstrate the presence of an extensive coiled-coil region between residues 463 and 692 of Rab11-FIP3, which exists as a dimer in solution and is critical to support its function on the ERC. Together, these data indicate that Rab11-FIP3 is necessary for the structural integrity of the pericentrosomal ERC.
authors: Molecular Cell Biology Laboratory, Department of Biochemistry, Biosciences Institute, University College Cork, Cork, Ireland.
authors: Conor P
authors: CP
authors: Horgan
authors: Arkadiusz
authors: A
authors: Oleksy
authors: Alexander V
authors: AV
authors: Zhdanov
authors: Patrick Y
authors: PY
authors: Lall
authors: Ian J
authors: IJ
authors: White
authors: Amir R
authors: AR
authors: Khan
authors: Clare E
authors: CE
authors: Futter
authors: John G
authors: JG
authors: McCaffrey
authors: Mary W
authors: MW
authors: McCaffrey
mesh_terms: D002352:Carrier Proteins / Q000502:physiology*; D045744:Cell Line, Tumor; D018385:Centrosome / Q000502:physiology; D011992:Endosomes / Q000737:chemistry / Q000502:physiology*; D006801:Humans
chemical_list: D002352:Carrier Proteins; C439063:RAB11FIP3 protein, human