Myosin VI regulates endocytosis of the cystic fibrosis transmembrane conductance regulator

被引:65
作者
Swiatecka-Urban, A [1 ]
Boyd, C
Coutermarsh, B
Karlson, KH
Barnaby, R
Aschenbrenner, L
Langford, GM
Hasson, T
Stanton, BA
机构
[1] Dartmouth Coll Sch Med, Dept Physiol, Hanover, NH 03755 USA
[2] Univ Calif San Diego, Sect Cell & Dev Biol, La Jolla, CA 92093 USA
[3] Dartmouth Coll, Dept Biol Sci, Hanover, NH 03755 USA
关键词
D O I
10.1074/jbc.M403141200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The cystic fibrosis transmembrane conductance regulator ( CFTR) is a cyclic AMP-regulated Cl- channel expressed in the apical plasma membrane in fluid-transporting epithelia. Although CFTR is rapidly endocytosed from the apical membrane of polarized epithelial cells and efficiently recycled back to the plasma membrane, little is known about the molecular mechanisms regulating CFTR endocytosis and endocytic recycling. Myosin VI, an actin-dependent, minus-end directed mechanoenzyme, has been implicated in clathrin-mediated endocytosis in epithelial cells. The goal of this study was to determine whether myosin VI regulates CFTR endocytosis. Endogenous, apical membrane CFTR in polarized human airway epithelial cells (Calu-3) formed a complex with myosin VI, the myosin VI adaptor protein Disabled 2 (Dab2), and clathrin. The tail domain of myosin VI, a dominant-negative recombinant fragment, displaced endogenous myosin VI from interacting with Dab2 and CFTR and increased the expression of CFTR in the plasma membrane by reducing CFTR endocytosis. However, the myosin VI tail fragment had no effect on the recycling of endocytosed CFTR or on fluid-phase endocytosis. CFTR endocytosis was decreased by cytochalasin D, an actin-filament depolymerizing agent. Taken together, these data indicate that myosin VI and Dab2 facilitate CFTR endocytosis by a mechanism that requires actin filaments.
引用
收藏
页码:38025 / 38031
页数:7
相关论文
共 68 条
[61]   Unconventional myosins, actin dynamics and endocytosis: A menage a trois? [J].
Soldati, T .
TRAFFIC, 2003, 4 (06) :358-366
[62]   PDZ domain interaction controls the endocytic recycling of the cystic fibrosis transmembrane conductance regulator [J].
Swiatecka-Urban, A ;
Duhaime, M ;
Coutermarsh, B ;
Karlson, KH ;
Collawn, J ;
Milewski, M ;
Cutting, GR ;
Guggino, WB ;
Langford, G ;
Stanton, BA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (42) :40099-40105
[63]  
Warren RA, 1997, J BIOL CHEM, V272, P2116
[64]   Distinct saturable pathways for the endocytosis of different tyrosine motifs [J].
Warren, RA ;
Green, FA ;
Stenberg, PE ;
Enns, CA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (27) :17056-17063
[65]   SUBCELLULAR-LOCALIZATION OF CFTR TO ENDOSOMES IN A DUCTAL EPITHELIUM [J].
WEBSTER, P ;
VANACORE, L ;
NAIRN, AC ;
MARINO, CR .
AMERICAN JOURNAL OF PHYSIOLOGY, 1994, 267 (02) :C340-C348
[66]   μ2 binding directs the cystic fibrosis transmembrane conductance regulator to the clathrin-mediated endocytic pathway [J].
Weixel, KM ;
Bradbury, NA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (49) :46251-46259
[67]   Myosin VI is an actin-based motor that moves backwards [J].
Wells, AL ;
Lin, AW ;
Chen, LQ ;
Safer, D ;
Cain, SM ;
Hasson, T ;
Carragher, BI ;
Milligan, RA ;
Sweeney, HL .
NATURE, 1999, 401 (6752) :505-508
[68]   MOLECULAR MECHANISMS OF CFTR CHLORIDE CHANNEL DYSFUNCTION IN CYSTIC-FIBROSIS [J].
WELSH, MJ ;
SMITH, AE .
CELL, 1993, 73 (07) :1251-1254