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J. Biol. Chem., Vol. 266, Issue 1, 143-154, Jan, 1991
PM Sullivan, P Petrusz, C Szpirer and DR Joseph
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.
Alternative processing of androgen-binding protein RNA transcripts in fetal rat liver. Identification of a transcript formed by trans splicing
Department of Biology, University of North Carolina, Chapel Hill 27599.
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