Alternative processing of androgen-binding protein RNA transcripts in fetal rat liver. Identification of a transcript...

Androgens and their nuclear receptor regulate genes necessary for development of the male phenotype, a process that is thought to be modulated by extracellular androgen carrier proteins. Two of these carrier proteins, testicular androgen-binding protein (ABP) and plasma sex hormone-binding globulin (SHBG), are encoded by the same gene, but differ in glycosylation and possibly amino acid sequence. To investigate ABP-SHBG gene expression in fetal rat liver, we analyzed RNA transcripts and expressed protein. These studies demonstrated a transient expression of ABP in hepatocytes during the time of testosterone-dependent differentiation of the Wolffian duct. Analysis of cDNA clones derived from fetal rat liver cDNA libraries identified two cDNAs encoded by the ABP-SHBG gene that represented alternatively spliced RNAs. One cDNA had an alternate exon 1, suggesting the function of another promoter in fetal liver. This cDNA also lacked testicular exon 6 DNA, an alteration that implicates the encoded protein in regulatory functions. The other cDNA represented a fused transcript of the ABP-SHBG gene (exons 1-5) and the histidine decarboxylase (HDC) gene, encoding a Mr 98,000 precursor protein. The two domains were joined at splice junctions of the ABP-SHBG and HDC genes, which were localized to rat chromosomes 10 and 3, respectively. Our results indicate that the joining of the two domains was by a trans (donor and acceptor)-splicing mechanism. Data from Northern hybridization experiments suggest the fusion transcript is present in fetal liver RNA. Polymerase chain reaction experiments with fetal liver cDNA further support the existence of an ABP-HDC fusion transcript, as well as the other alternate mRNA. Moreover, a Mr 93,000 immunoreactive protein was transiently expressed in fetal liver during the time of ABP and HDC gene expression. Expression of the fusion cDNA in COS cells yielded HDC activity and the predicted size protein (Mr = 93,000) on Western immunoblots.