Distinct pathways for basolateral targeting of membrane and secretory proteins in polarized epithelial cells.
Polarized epithelial cells target distinct sets of membrane and secretory proteins to their apical and basolateral domains. Here we examine whether constitutively secreted and membrane proteins that are bound for the same domain share the same carrier vesicles. To address the issue, differential effects of microtubule depolymerization on basolateral protein targeting in the polarized Madin-Darby canine kidney II cell line were studied. We find that the basolateral insertion of the active, ouabain-binding Na+,K(+)-ATPase and of a set of very late antigen integrins is little affected by microtubule disruption. Under equivalent conditions, the basolateral secretion of the basement membrane protein laminin is strongly suppressed. More specifically, it is demonstrated that microtubules are involved in targeting laminin, but not integrins, from the compartment related to the accumulation of newly synthesized proteins at 20 degrees C (trans-Golgi network) to the basolateral domain. Our study also reveals that laminin associated with basolateral binding sites interacts with those sites only secondarily to secretion. The data provide evidence for a branch in the basolateral targeting pathway, with secreted and membrane proteins loaded into distinct carrier vesicles.
Boll
W
W
Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510-8002.
Partin
J S
JS
Katz
A I
AI
Caplan
M J
MJ
Jamieson
J D
JD
D000818:Animals; D001485:Basement Membrane / Q000378:metabolism; D002451:Cell Compartmentation / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003078:Colchicine / Q000494:pharmacology; D004285:Dogs; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure*; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000378:metabolism; D007797:Laminin / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D008870:Microtubules / Q000187:drug effects / Q000502:physiology; D016029:Receptors, Very Late Antigen / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism
D007797:Laminin; D008565:Membrane Proteins; D016029:Receptors, Very Late Antigen; D014404:Tubulin; D000254:Sodium-Potassium-Exchanging ATPase; D003078:Colchicine