Identification and androgen-regulated expression of two major human glandular kallikrein-1 (hGK-1) mRNA species.
The screening of an oligo(dT)-primed prostate cDNA library with a human glandular kallikrein-1 (hGK-1) genomic DNA fragment resulted in the isolation of two different hGK-1 cDNAs. A 1.2 kb cDNA (pGK-1) contains an open reading frame of 510 bp, encoding the major part of the previously predicted hGK-1 protein (Schedlich et al. (1987) DNA 6, 429-437). This cDNA contains a 3'-untranslated region of 677 nucleotides and terminates in a poly(A) stretch, preceded by the canonical AATAAA polyadenylation signal. A second cDNA (pGK-10A), with a size of 1.5 kb, contains an open reading frame of 669 nucleotides preceded by 16 nucleotides of the 5'-untranslated region. pGK-10A differs from pGK-1 by the presence of an additional 37 bp fragment, interrupting the protein coding region of hGK-1, which results from the use of an alternative splice donor site of intron IV of the hGK-1 gene. The mature protein (excluding presumed pre- and propeptides) as deduced from the pGK-10A cDNA sequence, has a size of 199 amino acids and differs at the COOH-terminus from the 237 amino acid hGK-1 protein. The alternatively spliced mRNA comprises approximately 20% of the hGK-1 transcripts, as deduced from analysis of mRNA from prostate cells by PCR amplification of specific fragments. The regulation of hGK-1 mRNA expression was studied in different human prostate tumors and cell lines by Northern blotting, using a hGK-1-specific oligonucleotide probe. A high level of hGK-1 expression was found in the androgen-dependent tumors PC 82 and PC EW. hGK-1 mRNA was also present in the androgen-sensitive LNCaP cell line, but undetectable in the androgen-insensitive prostate tumors PC 133, PC 135 and the PC 3 cell line. In LNCaP cells, the expression of hGK-1 mRNA was strongly induced by androgens. Regulation of expression of the closely related prostate-specific antigen (PA) gene showed a similar pattern.
Riegman
P H
PH
Department of Pathology, Erasmus University, Rotterdam, The Netherlands.
Vlietstra
R J
RJ
van der Korput
H A
HA
Romijn
J C
JC
Trapman
J
J
D000595:Amino Acid Sequence; D000728:Androgens / Q000502:physiology*; D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D004247:DNA; D005786:Gene Expression Regulation; D006801:Humans; D007610:Kallikreins / Q000096:biosynthesis / Q000235:genetics*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D017430:Prostate-Specific Antigen; D012333:RNA, Messenger / Q000096:biosynthesis*; D015183:Restriction Mapping; D014407:Tumor Cells, Cultured
D000728:Androgens; D000951:Antigens, Neoplasm; D012333:RNA, Messenger; D004247:DNA; D007610:Kallikreins; D017430:Prostate-Specific Antigen