A novel alternative spliced chondrolectin isoform lacking the transmembrane domain is expressed during T cell maturation

Chondrolectin (CHODL) is a novel type I transmembrane protein containing one carbohydrate recognition domain (CRD) of C-type lectins. Recently, data base searching revealed a variant of CHODL (AK022689) with a different 5' leader sequence derived from a new putative upstream alternative promoter (P2). The P2 promoter gives rise to at least three additional alternatively spliced isoforms, designated as CHODLf, CHODLf Delta E, and CHODL Delta E. Of all variants, the alternative exon E-splicing isoforms (CHODLf Delta E/CHODL Delta E) are expressed exclusively in the T lymphocyte lineage and are regulated during T lymphopoiesis. Peripheral T lymphocytes demonstrated a unique exon E-splicing pattern in comparison with end maturation stage thymocytes, suggesting its association with the post-thymic maturation of T cells. Since exon E encodes the transmembrane domain of CHODL, the exon E-skipping variant results in a non-transmembrane domain-containing isoform (CHODLf Delta E/CHODL Delta E) terminating in the QDEL sequence, thus suggesting different functional attributes of CHODL isoforms during the development of T cells. Double label immunofluoresence experiments demonstrated that the transmembrane-containing isoform (CHODLf) colocalizes with rBet1 to the endoplasmic reticulum-Golgi apparatus. In summary, this study describes the molecular characterization of novel members of the chondrolectin family associated with T cell maturation and a subcellular localization of CHODLf in the endoplasmic reticulum-Golgi apparatus.     

Differential modulation of cadherin-mediated cell-cell adhesion by platelet endothelial cell adhesion molecule-1 isoforms through activation of extracellular regulated kinases

The role of platelet endothelial cell adhesion molecule-1 (PECAM-1) in endothelial cell-cell interactions and its contribution to cadherin-mediated cell adhesion are poorly understood. Such studies have been difficult because all known endothelial cells express PECAM-1. We have used Madin-Darby canine kidney (MDCK) cells as a model system in which to evaluate the role of PECAM-1 isoforms that differ in their cytoplasmic domains in cell-cell interactions. MDCK cells lack endogenous PECAM-1 but form cell-cell junctions similar to those of endothelial cells, in which PECAM-1 is concentrated. MDCK cells were transfected with two isoforms of murine PECAM-1, Delta15 and Delta14&15, the predominant isoforms expressed in vivo. Expression of the Delta15 isoform resulted in apparent dedifferentiation of MDCK cells concomitant with the loss of adherens junctions, down-regulation of E-cadherin, alpha- and beta-catenin expression, and sustained activation of extracellular regulated kinases. The Delta15 isoform was not concentrated at cell-cell contacts. In contrast, the Delta14&15 isoform localized to sites of cell-cell contact and had no effect on MDCK cell morphology, cadherin/catenin expression, or extracellular regulated kinase activity. Thus, the presence of exon 14 in the cytoplasmic domain of PECAM-1 has dramatic effects on the ability of cells to maintain adherens junctions and an epithelial phenotype. Therefore, changes in the expression of exon 14 containing PECAM-1 isoforms, which we have observed during development, may have profound functional consequences.