Isolation and characterization of mouse probasin: An androgen-regulated protein specifically expressed in the differentiated prostate.
BACKGROUND
The development and growth of the prostate gland is regulated, in part, by a variety of steroid and polypeptide growth-factor hormones. As a consequence of hormone action, the prostate gland will produce a number of tissue-restricted gene products. Characterization of the regulation, expression, and function of genes encoding prostate-specific proteins is critical to our understanding of prostate biology. Probasin is a prostate-specific gene originally isolated from the rat and has been exploited as a marker of prostate differentiation and to elucidate androgen action. Furthermore, a number of transgenic mouse models of prostate cancer have been established based on the regulatory elements derived from the rat probasin gene. In this report, we describe the isolation and characterization of the mouse probasin ortholog to further facilitate studies related to hormone action in the prostate and the generation and characterization of novel autochthonous models of prostate cancer.


METHODS
Mouse probasin cDNA was isolated from a phage library, and the DNA sequence was determined. The predicted protein sequence was used to generate specific oligonucleotide primers and antibodies. Probasin protein and RNA expression were examined by immunobloting, immunohistochemistry, and RT-PCR, in normal mouse prostate tissue and tumor tissues derived from the autochthonous "transgenic adenocarcinoma of the mouse prostate" (TRAMP) model. Regulation of probasin expression in response to surgical castration and hormone supplementation was also characterized.


RESULTS
Several points of evolutionary sequence conservation were identified between mouse and rat probasin, especially in the 3' untranslated region. Specific polyclonal antibodies were generated to peptide fragments, and the temporal and spatial pattern of probasin expression was examined. The expression of probasin was primarily localized to the apical membrane of differentiated secretory epithelium. Probasin mRNA and protein were absent from the poorly differentiated tissue of TRAMP tumors. Probasin was found to be androgen-regulated. In contrast to data from studies on rat probasin, no postcastration rebound of mouse probasin mRNA was observed.


CONCLUSIONS
Probasin is a marker of differentiation and androgen action in the mouse prostate, and strong sequence conservation between mouse and rat probasin supports an essential role for this gene in the biology of the prostate gland. Isolation and characterization of mouse probasin will facilitate further development and analysis of autochthonous mouse models of prostate cancer.
Johnson
M A
MA
Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
Hernandez
I
I
Wei
Y
Y
Greenberg
N
N
D000595:Amino Acid Sequence; D000727:Androgen-Binding Protein / Q000302:isolation & purification*; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002454:Cell Differentiation; D018076:DNA, Complementary / Q000737:chemistry; D007106:Immune Sera; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011467:Prostate / Q000378:metabolism*; D011817:Rabbits; D051381:Rats; D011503:Transglutaminases / Q000378:metabolism*
D000727:Androgen-Binding Protein; D018076:DNA, Complementary; D007106:Immune Sera; C027519:probasin; D011503:Transglutaminases