Cdc42-dependent modulation of tight junctions and membrane protein traffic in polarized Madin-Darby canine kidney cells.
Polarized epithelial cells maintain the asymmetric composition of their apical and basolateral membrane domains by at least two different processes. These include the regulated trafficking of macromolecules from the biosynthetic and endocytic pathway to the appropriate membrane domain and the ability of the tight junction to prevent free mixing of membrane domain-specific proteins and lipids. Cdc42, a Rho family GTPase, is known to govern cellular polarity and membrane traffic in several cell types. We examined whether this protein regulated tight junction function in Madin-Darby canine kidney cells and pathways that direct proteins to the apical and basolateral surface of these cells. We used Madin-Darby canine kidney cells that expressed dominant-active or dominant-negative mutants of Cdc42 under the control of a tetracycline-repressible system. Here we report that expression of dominant-active Cdc42V12 or dominant-negative Cdc42N17 altered tight junction function. Expression of Cdc42V12 slowed endocytic and biosynthetic traffic, and expression of Cdc42N17 slowed apical endocytosis and basolateral to apical transcytosis but stimulated biosynthetic traffic. These results indicate that Cdc42 may modulate multiple cellular pathways required for the maintenance of epithelial cell polarity.
Rojas
R
R
Renal-Electrolyte Division of the Department of Medicine, Laboratory of Epithelial Biology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
Ruiz
W G
WG
Leung
S M
SM
Jou
T S
TS
Apodaca
G
G
D000199:Actins / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D016764:Cell Polarity; D003599:Cytoskeleton / Q000378:metabolism; D004285:Dogs; D017097:Electric Impedance; D004705:Endocytosis / Q000502:physiology; D004815:Epidermal Growth Factor / Q000378:metabolism; D000077614:Golgi Matrix Proteins; D015151:Immunoblotting; D007070:Immunoglobulin A / Q000378:metabolism; D007444:Inulin / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D062793:Occludin; D010750:Phosphoproteins / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D019108:Tight Junctions / Q000378:metabolism* / Q000648:ultrastructure; D062826:Zonula Occludens-1 Protein; D020764:cdc42 GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
D000199:Actins; D000077614:Golgi Matrix Proteins; D007070:Immunoglobulin A; D008565:Membrane Proteins; D062793:Occludin; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D062826:Zonula Occludens-1 Protein; C087574:macrogolgin; D004815:Epidermal Growth Factor; D007444:Inulin; D020764:cdc42 GTP-Binding Protein