Primary structure and androgen regulation of a 20-kilodalton protein specific to rat ventral prostate.
Nuclear and cytosolic forms of a 20-kdalton rat ventral prostate protein were purified and partially sequenced from their N-termini. Isolated nuclei were treated with micrococcal nuclease and extracted in 0.6 M NaCl, and proteins were separated by affinity chromatography on Matrex gel green A, ammonium sulfate fractionation, and fast protein liquid chromatography on Superose 12. The 43 amino acid N-terminal sequence of the nuclear 20-kdalton protein was identical with the cytosolic protein except it lacked 7 N-terminal amino acids present in the cytosolic form. The DNA sequence of a full-length complementary DNA clone isolated from a ventral prostate gt11 library extended the N-terminal sequence of the cytosolic form by an additional nine amino acids from the predicted initiation methionine. The cDNA included the nucleotide sequence for the 43 amino acid N-terminal sequence of the purified 20-kdalton protein and predicted molecular weights of 16,686, 17,521, and 18,650, respectively, for the nuclear, cytoplasmic, and nonprocessed proteins. Northern blot analyses of reproductive tract tissue RNAs using the 20-kdalton protein cDNA as probe revealed a single mRNA species of 0.92 kb detectable only in extracts of rat ventral prostate. Expression of the 0.92-kb mRNA was androgen dependent since the mRNA was undetectable in extracts obtained 4 days after castration and was restored 16 h after restimulation with androgen.
Ho
K C
KC
Laboratories for Reproductive Biology, University of North Carolina, Chapel Hill 27599.
Snoek
R
R
Quarmby
V
V
Viskochil
D H
DH
Rennie
P S
PS
Wilson
E M
EM
French
F S
FS
Bruchovsky
N
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D009919:Orchiectomy; D011467:Prostate / Q000187:drug effects / Q000378:metabolism*; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification*; D012313:RNA / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D051381:Rats; D011919:Rats, Inbred Strains; D015183:Restriction Mapping; D013739:Testosterone / Q000494:pharmacology*
D009687:Nuclear Proteins; D011506:Proteins; D013739:Testosterone; D012313:RNA; D004247:DNA