CHARACTERIZATION OF THE HAMSTER DDT-1 CELL AFGF/HGBF-I GENE AND CDNA AND ITS MODULATION BY STEROIDS

Syrian hamster DDT‐1 cells are derived from smooth muscle of the ductus deferens. DDT‐1 cell growth is increased by the addition of testosterone (T). Acidic fibroblast growth factor (aFGF) or basic fibroblast growth factor (bFGF) also known as heparin binding growth factor I and II (HBGF‐I and HBGF‐II) can replace T in the stimulation of growth in these cells. This phenomenon is correlated with testosterone's ability to elevate aFGF/HBGF‐I mRNA. The increase steady‐state levels of aFGF/HBGF‐I mRNA were documented by northern blots and by in situ hybridization. Using a 520 bp human aFGF/HBGF‐I cDNA probe, a genomic clone with a 38 kb DNA insert was isolated from a cosmid library. By restriction enzyme analysis and southern hybridization, it was determined that there are three coding exons. DNA sequence analysis showed all of the coding region and 3′ noncoding sequences were on this clone. A 5′ noncoding exon not in the 38 kb insert is indicated, based on the cDNA sequences and genomic sequences of aFGF/HBGF‐I's from hamster DDT‐1 cells and several other species. The cDNA for hamster aFGF/HBGF‐I was isolated from a DDT‐1 lambda gt11 library and sequenced. Comparison of the coding region of aFGF/HBGF‐I from four species shows a >90% conservation of amino acid sequence. Copyright © 1990 Wiley‐Liss, Inc.