Abstract
Transforming growth factor β4 (TGF-β4) 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-β4. By using a GC-rich PCR system in a modified 5 ′RACE methodology we generated the 5 ′-end of cDNA sequence encoding the TGF-β4 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-β4 precursor protein was established from CHO-K1 cells. Acid-activated mature TGF-β4 inhibited the growth of mink lung epithelial (Mv1Lu) cell line. TGF-β4 also stimulated the expression of type I procollagen and enhanced heat shock protein 47 (Hsp47) expression in chicken tendon fibroblasts. Hsp47 expression by TGFβ4 is likely regulated through activation of heat shock transcription factor 1 (HSF1). Because the presence of TGF-β1 has not been documented in avian cells and our data show that TGF-β4 elicits biological activities in chicken tendon cells, which closely parallel that of TGF-β1, we propose that TGF-β4 plays roles in avian species similar to what TGF-β1 plays in mammalian species.
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