Novel non-TGF-β-binding splice variant of LTBP-4 in human cells and tissues provides means to decrease TGF-β deposition

Small latent TGF-β consists of latency associated peptide (LAP) bound to the 25 kDa TGF-β by noncovalent interactions. Small latent TGF-β is secreted from cells and deposited into the extracellular matrix as covalent complexes with its binding proteins, LTBPs. Four LTBPs have been molecularly cloned and their structures contain repetitive sequences. The 3rd 8-Cys repeats of LTBP-1, -3 and -4 are able to associate with small latent TGF-β. We analyzed by RT-PCR the expression of LTBPs 1-4 in a panel of cultured human cell lines including fibroblasts of different origin, endothelial cells and immortalized keratinocytes. LTBPs were expressed in an overlapping manner, but differences in their expression levels were detected. SV-40 transformed human embryonic lung fibroblasts contained less of the mRNAs for the LTBPs, suggesting that malignant transformation leads to decrease in LTBP expression. A novel alternatively spliced form of LTBP-4 lacking the 3rd 8-Cys repeat (LTBP-4Δ8-Cys3rd) was identified. LTBP-4Δ8-Cys3rd does not bind TGF-β and it was found to be expressed in the same tissues as the full length LTBP-4. The exon-intron structure of LTBP-4 around the 3rd 8-Cys repeat was similar to those of LTBP-2 and -3. LTBP-4Δ8-Cys3rd was produced by alternative splicing over two exons. In addition, HL-60 promyelocytic leukemia cells expressed a splice variant lacking only one exon of this region. The expression of the non-TGF-β-binding variant of LTBP-4 may be important for the regulation of TGF-β deposition in tissues. Since LTBPs are a part of the extracellular matrix microfibrils, the LTBP-4Δ8-Cys3rd protein may also be involved in various structural functions not related to TGF-β signaling.