Reference : CLC-5 and KIF3B interact to facilitate CLC-5 plasma membrane expression, endocytosis,...
Scientific journals : Article
Life sciences : Genetics & genetic processes
CLC-5 and KIF3B interact to facilitate CLC-5 plasma membrane expression, endocytosis, and microtubular transport: relevance to pathophysiology of Dent's disease.
Reed, Anita A. C. [> >]
Loh, Nellie Y. [> >]
Terryn, Sara [> >]
Lippiat, Jonathan D. [> >]
Partridge, Chris [> >]
Galvanovskis, Juris [> >]
Williams, Sian E. [> >]
JOURET, François mailto [Centre Hospitalier Universitaire de Liège - CHU > > Néphrologie >]
Wu, Fiona T. F. [> >]
Courtoy, Pierre J. [> >]
Nesbit, M. Andrew [> >]
Rorsman, Patrik [> >]
Devuyst, Olivier [> >]
Ashcroft, Frances M. [> >]
Thakker, Rajesh V. [> >]
American Journal of Physiology - Renal Physiology
Yes (verified by ORBi)
United States
[en] Adult ; Albumins/metabolism ; Animals ; COS Cells ; Cell Line ; Cercopithecus aethiops ; Chloride Channels/metabolism/physiology ; DNA, Complementary ; Down-Regulation ; Drug Interactions ; Electric Conductivity ; Endocytosis/physiology ; Gene Library ; Genetic Diseases, X-Linked/physiopathology ; Humans ; Kidney/cytology/metabolism ; Kidney Diseases/physiopathology ; Kidney Tubules, Proximal/cytology/metabolism ; Kinesin/metabolism ; Mice ; Mice, Knockout ; Microtubules/metabolism ; Protein Interaction Domains and Motifs ; Protein Transport ; Two-Hybrid System Techniques ; Up-Regulation
[en] Renal tubular reabsorption is important for extracellular fluid homeostasis and much of this occurs via the receptor-mediated endocytic pathway. This pathway is disrupted in Dent's disease, an X-linked renal tubular disorder that is characterized by low-molecular-weight proteinuria, hypercalciuria, nephrolithiasis, and renal failure. Dent's disease is due to mutations of CLC-5, a chloride/proton antiporter, expressed in endosomes and apical membranes of renal tubules. Loss of CLC-5 function alters receptor-mediated endocytosis and trafficking of megalin and cubilin, although the underlying mechanisms remain to be elucidated. Here, we report that CLC-5 interacts with kinesin family member 3B (KIF3B), a heterotrimeric motor protein that facilitates fast anterograde translocation of membranous organelles. Using yeast two-hybrid, glutathione-S-transferase pull-down and coimmunoprecipitation assays, the COOH terminus of CLC-5 and the coiled-coil and globular domains of KIF3B were shown to interact. This was confirmed in vivo by endogenous coimmunoprecipitation of CLC-5 and KIF3B and codistribution with endosomal markers in mouse kidney fractions. Confocal live cell imaging in kidney cells further demonstrated association of CLC-5 and KIF3B, and transport of CLC-5-containing vesicles along KIF3B microtubules. KIF3B overexpression and underexpression, using siRNA, had reciprocal effects on whole cell chloride current amplitudes, CLC-5 cell surface expression, and endocytosis of albumin and transferrin. Clcn5(Y/-) mouse kidneys and isolated proximal tubular polarized cells showed increased KIF3B expression, whose effects on albumin endocytosis were dependent on CLC-5 expression. Thus, the CLC-5 and KIF3B interaction is important for CLC-5 plasma membrane expression and for facilitating endocytosis and microtubular transport in the kidney.

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