Monoclonal antibodies in the analysis of fibronectin isoforms generated by alternative splicing of mRNA precursors in normal and transformed human cells.
Recent results showing that a single fibronectin gene can give rise to several different mRNAs by alternative splicing have offered an explanation for fibronectin polymorphism. Here we report on monoclonal antibodies that show specificity for a fibronectin segment (ED) that can be included or omitted from the molecule depending on the pattern of splicing of the mRNA precursors. Using these monoclonals, we have quantitatively analyzed the expression of the ED sequence in human fibronectin from different sources. The results demonstrated that, at the protein level, the ED segment is not expressed in plasma fibronectin and that, in fibronectin from the tissue culture medium of tumor-derived or simian virus-40-transformed human cells, the percentage of fibronectin molecules containing the ED segment is about 10 times higher than in fibronectin from normal human fibroblasts. These results suggest that in malignant cells the mechanisms that regulate the splicing of mRNA precursors are altered.
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D005353:Fibronectins / Q000032:analysis / Q000235:genetics*; D006801:Humans; D008970:Molecular Weight; D009369:Neoplasms; D009694:Nucleic Acid Precursors / Q000235:genetics*; D010446:Peptide Fragments / Q000032:analysis; D012322:RNA Precursors; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics*
D000911:Antibodies, Monoclonal; D005353:Fibronectins; D009694:Nucleic Acid Precursors; D010446:Peptide Fragments; D012322:RNA Precursors; D012333:RNA, Messenger