Effect of nocodazole on vesicular traffic to the apical and basolateral surfaces of polarized MDCK cells

A polarized cell, to maintain distinct basolateral and apical membrane domains, must tightly regulate vesicular traffic terminating at either membrane domain. In this study we have examined the extent to which microtubules regulate such traffic in polarized cells. Using the polymeric immunoglobulin receptor expressed in polarized MDCK cells, we have examined the effects of nocodazole, a microtubule-disrupting agent, on three pathways that deliver proteins to the apical surface and two pathways that deliver proteins to the basolateral surface. The biosynthetic and transcytotic pathways to the apical surface are dramatically altered by nocodazole in that a portion of the protein traffic on each of these two pathways is misdirected to the basolateral surface. The apical recycling pathway is slowed in the presence of nocodazole but targeting is not disrupted. In contrast, the biosynthetic and recycling pathways to the basolateral surface are less affected by nocodazole and therefore appear to be more resistant to microtubule disruption

Deletions in the cytoplasmic domain of the polymeric immunoglobulin receptor differentially affect endocytotic rate and postendocytotic traffic

We have examined the function of the cytoplasmic domain of the polymeric immunoglobulin receptor (pIg-R) by producing two separate deletions in the cytoplasmic domain of the pIg-R, expressing the mutant receptors in polarized MDCK cells, and analyzing each for their effects on receptor and ligand traffic. Deletion of the C-terminal 30 amino acids (726-755) reduces the rate of internalization of receptor-bound ligand from the basolateral surface. However, this mutation has no effect on delivery of receptor from the Golgi to the basolateral surface or the post-endocytotic traffic of receptor and ligand. Mutation of a tyrosine at position 734 to serine produces a receptor with a similar phenotype. If residues 670-707 are deleted from the middle of the cytoplasmic domain, both basolateral delivery and internalization are unaffected. However, unlike wild type, after endocytosis from the basolateral surface, both receptor and ligand are largely degraded. We reported previously that deletion of the entire cytoplasmic domain prevents the basolateral delivery of newly synthesized receptor (Mostov, K.E., de Bruyn Kops, A., and Deitcher, D.L. (1986) Cell 47, 359-364). In contrast, the mutants reported here are delivered to the basolateral surface, suggesting that only residues 653-669 and/or 708-725 are necessary for basolateral delivery. Thus, different deletions in the cytoplasmic domain of the pIg-R can produce mutant receptors which alter different aspects of receptor traffic