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Over-expression of Rififylin, a new RING finger and FYVE-like domain-containing protein, inhibits recycling from the endocytic recycling compartment.

By Franck Coumailleau, Vincent Das, Andres Alcover, Graça Raposo, Sandrine Vandormael-Pournin, Stéphanie Le Bras, Patricia Baldacci, Alice Dautry-Varsat, Charles Babinet and Michel Cohen-Tannoudji

Abstract

International audienceEndocytosed membrane components are recycled to the cell surface either directly from early/sorting endosomes or after going through the endocytic recycling compartment (ERC). Studying recycling mechanisms is difficult, in part due to the fact that specific tools to inhibit this process are scarce. In this study, we have characterized a novel widely expressed protein, named Rififylin (Rffl) for RING Finger and FYVE-like domain-containing protein, that, when overexpressed in HeLa cells, induced the condensation of transferrin receptor-, Rab5-, and Rab11-positive recycling tubulovesicular membranes in the perinuclear region. Internalized transferrin was able to access these condensed endosomes but its exit from this compartment was delayed. Using deletion mutants, we show that the carboxy-terminal RING finger of Rffl is dispensable for its action. In contrast, the amino-terminal domain of Rffl, which shows similarities with the phosphatidylinositol-3-phosphate-binding FYVE finger, is critical for the recruitment of Rffl to recycling endocytic membranes and for the inhibition of recycling, albeit in a manner that is independent of PtdIns(3)-kinase activity. Rffl overexpression represents a novel means to inhibit recycling that will help to understand the mechanisms involved in recycling from the ERC to the plasma membrane

Topics: MESH: Amino Acid Sequence, MESH: Animals, MESH: Mutation, MESH: Protein Structure, Tertiary, MESH: Proteins, MESH: Recombinant Fusion Proteins, MESH: Sequence Alignment, MESH: Transferrin, MESH: Zinc Fingers, MESH: Base Sequence, MESH: Endocytosis, MESH: Endosomes, MESH: Hela Cells, MESH: Humans, MESH: Intracellular Membranes, MESH: Mice, MESH: Molecular Sequence Data, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology
Publisher: 'American Society for Cell Biology (ASCB)'
Year: 2004
DOI identifier: 10.1091/mbc.E04-04-0274
OAI identifier: oai:HAL:pasteur-00166952v1
Provided by: HAL-Pasteur
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