R K Chiu | PubMed Health

PubMed1995

Adhesive interactions between alternatively spliced CD44 isoforms.

Alternative splicing of a series of 10 contiguous exons present within the CD44 gene can generate a large number of differentially expressed CD44 isoforms that contain additional peptide sequences of varying length inserted into a single site within the extracellular domain of the molecule proximal to the membrane spanning domain. Although distinct functions have been ascribed to certain of these isoforms, the effect of particular inserted domains on the ligand-binding specificity of the CD44 molecule remains unclear. In the present study, we demonstrate that while CD44H, the major CD44 isoform expressed on resting hemopoietic cells, and CD44R1, an alternatively spliced isoform present on transformed epithelial cells and certain activated and/or malignant hemopoietic cell types, can both bind avidly to hyaluronan, only CD44R1 can promote homotypic cellular aggregation when expressed in the CD44-negative murine lymphoma cell line TIL1. Experiments in which TIL1 cells transduced with different CD44 isoforms were tested for their ability to adhere to one another or to COS7 cells transfected with CD44R1, indicated that CD44R1 can recognize and bind a common determinant present on both CD44H and CD44R1. Monoclonal antibody blocking studies suggest further, that the determinant recognized by CD44R1 is located in a region of the CD44 molecule distinct from that involved in hyaluronan binding.

Alternative Splicing↗

PubMed1994

Ligand binding specificity of alternatively spliced CD44 isoforms. Recognition and binding of hyaluronan by CD44R1.

CD44 species of widely differing molecular mass have been identified on various normal and/or transformed cells. Recent studies have demonstrated that much of this heterogeneity is produced as a result of the alternative splicing of a series of 10 exons present within the CD44 gene generating a large number of CD44 isoforms containing additional peptide sequences of varying length inserted into a single site within the extracellular domain of the molecule. At present, the effect of such insertions on the ligand binding specificity of CD44 remains unclear. CD44H, the major CD44 isoform expressed by most resting cell types, has been shown to function as a receptor for the glycosaminoglycan hyaluronan. In contrast, CD44E, the major isoform expressed by the colon carcinoma cell line HT29, which contains a 132-amino acid insert, is unable to recognize and bind this ligand. In the present study we demonstrate that CD44R1, an isoform isolated from the myelomonocytic cell line KG1a, that differs from CD44E by just 3 amino acid substitutions, is fully capable of mediating the attachment of transfected COS7 cells to hyaluronan-coated plastic. In order to confirm that such binding was directly mediated by the introduced CD44 species, chimeric proteins containing the entire extracellular domain of CD44H or CD44R1 fused in-frame to human bone/liver/kidney alkaline phosphatase were prepared and tested for their ability to bind hyaluronan-coated plastic. Both fusion proteins bound equally well to hyaluronan and in each case their attachment could be readily inhibited by monoclonal antibodies directed against the hyaluronan-binding domain of CD44. These data indicate that the 132-amino acid insert present within the extracellular domain of CD44R1 does not interfere with the hyaluronan binding function of the molecule. Since CD44E contains an identically sized insert but is unable to bind hyaluronan, it is likely that mutation of one or more of the 3 amino acid residues that differ between CD44E and CD44R1 is responsible for the altered functional activity of this particular molecule.

Alternative Splicing↗