Role of Hrb in clathrin dependent endocytosis. - CNRS - Centre national de la recherche scientifique Accéder directement au contenu
Article Dans Une Revue Journal of Biological Chemistry Année : 2008

Role of Hrb in clathrin dependent endocytosis.

Lydia Danglot
Thierry Galli

Résumé

HIV Rev binding protein (Hrb), also called human Rev Interacting Protein (hRIP) or Rev/Rex Activation domain-Binding (RAB) is a partner of the tyrosine kinase substrate Eps15 and it has been recovered in the AP2 interactome. Eps15 and AP2 are involved in endocytosis, but the function of Hrb in this process is still unknown. Here, we have identified Hrb as a partner of the vesicular SNARE Tetanus neurotoxin Insensitive Vesicle-Associated Membrane Protein (TI-VAMP, also called VAMP7) in yeast two hybrid screens and using biochemical assays. In HeLa cells, Hrb localizes both in the nucleus and in the cytoplasm. In the cytoplasm, Hrb colocalizes with clathrin, AP2, Eps15 and transferrin receptor containing vesicles. We did not see significant colocalisation between Hrb and TI-VAMP in HeLa cells and partial colocalisation with GFP-TIVAMP in stably expessing MDCK cells. Nevertheless, using a pHLuorin-tagged TI-VAMP construct, we find that Hrb and TI-VAMP colocalize close to the plasma membrane after 5 minutes of anti-GFP antibody uptake. These results suggest that TI-VAMP and Hrb may interact only during the early stages of endocytosis. Furthermore, uptake experiments followed by fluorescence-activated cell sorting (FACS) show that the endocytosis of flurorescent transferrin and pHLuorin-TI-VAMP is strongly reduced in Hrb knock-down cells. Altogether, these results suggest that Hrb is involved in clathrin-dependent endocytosis and recruits TI-VAMP in this process.

Dates et versions

hal-00326921 , version 1 (06-10-2008)

Identifiants

Citer

Mathilde Chaineau, Lydia Danglot, Véronique Proux-Gillardeaux, Thierry Galli. Role of Hrb in clathrin dependent endocytosis.. Journal of Biological Chemistry, 2008, epub ahead of print. ⟨10.1074/jbc.M804587200⟩. ⟨hal-00326921⟩
36 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More