Cloning, expression, and characterization of chicken transforming growth factor beta 4

Transforming growth factor beta 4 (TGF-beta 4) is unique to avian species, though its roles in vivo have not yet been well established. In this paper we describe the expression and partial characterization of recombinant chicken TGF-beta 4. By using a GC-rich PCR system in a modified 5'RACE methodology we generated the 5'-end of cDNA sequence encoding the TGF-beta 4 precursor, which was in-frame cloned into pcDNA3.1/V5-His-TOPO and transfected into the Chinese hamster ovary cell line (CHO-K1). A cell line stably expressing TGF-beta 4 precursor protein was established from CHO-K1 cells. Acid-activated mature TGF-beta 4 inhibited the growth of mink lung epithelial (Mv1Lu) cell line. TGF-beta 4 also stimulated the expression of type I procollagen and enhanced heat shock protein 47 (Hsp47) expression in chicken tendon fibroblasts. Hsp47 expression by TGF beta 4 is likely regulated through activation of heat shock transcription factor 1 (HSF1). Because the presence of TGF-beta 1 has not been documented in avian cells and our data show that TGF-beta 4 elicits biological activities in chicken tendon cells, which closely parallel that of TGF-beta 1, we propose that TGF-beta 4 plays roles in avian species similar to what TGF-beta 1 plays in mammalian species.