Analysis of Articulation Between Clathrin and Retromer in Retrograde Sorting on Early Endosomes

Popoff, Vincent; Mardones, Gonzalo A.; Bai, Siau-Kun; Chambon, Valerie; Tenza, Daniele; Burgos, Patricia V.; Shi, Anbing; Benaroch, Philippe; Urbe, Sylvie; Lamaze, Christophe; Grant, Barth D.; Raposo, Graca; Johannes, Ludger

Abstract

Clathrin and retromer have key functions for retrograde trafficking between early endosomes and the trans-Golgi network (TGN). Previous studies on Shiga toxin suggested that these two coat complexes operate in a sequential manner. Here, we show that the curvature recognition subunit component sorting nexin 1 (SNX1) of retromer interacts with receptor-mediated endocytosis-8 (RME-8) protein, and that RME-8 and SNX1 colocalize on early endosomes together with a model cargo of the retrograde route, the receptor-binding B-subunit of Shiga toxin (STxB). RME-8 has previously been found to bind to the clathrin uncoating adenosine triphosphatase (ATPase) Hsc70, and we now report that depletion of RME-8 or Hsc70 affects retrograde trafficking at the early endosomes-TGN interface of STxB and the cation-independent mannose 6-phosphate receptor, an endogenous retrograde cargo protein. We also provide evidence that retromer interacts with the clathrin-binding protein hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs) not only via SNX1, as previously published (Chin Raynor MC, Wei X, Chen HQ, Li L. Hrs interacts with sorting nexin 1 and regulates degradation of epidermal growth factor receptor. J Biol Chem 2001;276:7069-7078), but also via the core complex component Vps35. Hrs codistributes at the ultrastructural level with STxB on early endosomes, and interfering with Hrs function using antibodies or mild overexpression inhibits retrograde transport. Our combined data suggest a model according to which the functions in retrograde sorting on early endosomes of SNX1/retromer and clathrin are articulated by RME-8, and possibly also by Hrs.

Más información

Título según WOS: ID WOS:000271714300013 Not found in local WOS DB
Título de la Revista: TRAFFIC
Volumen: 10
Número: 12
Editorial: Wiley
Fecha de publicación: 2009
Página de inicio: 1868
Página final: 1880
DOI:

10.1111/j.1600-0854.2009.00993.x

Notas: ISI