Transcriptional activation of the human prostatic acid phosphatase gene by NF-kappaB via a novel hexanucleotide-binding site.
Human prostatic acid phosphatase (PAcP) is a prostate epithelium-specific differentiation antigen. Cellular PAcP functions as a neutral protein tyrosine phosphatase and is involved in regulating androgen-promoted prostate cancer cell proliferation. Despite the fact that the promoter of the PAcP gene has been cloned, the transcriptional factors that regulate PAcP expression remain unidentified. This article describes our analyses of the promoter of the PAcP gene. Deletion analyses of the promoter sequence up to -4893 (-4893/+87) revealed that a 577 bp fragment (-1356/-779) represents the unique positive cis-active element in human prostate cancer cells but not in HeLa cervix carcinoma cells. Interestingly, the 577 bp fragment contains a non-consensus nuclear factor kappaB (NF-kappaB)-binding site that is required for NF-kappaB up-regulation in prostate cancer cells, while NF-kappaB failed to have the same effect in HeLa cells. Conversely, inhibition of the NF-kappaB pathway stopped p65 NF-kappaB activation of the p1356 promoter activity. Gel shift and mutation analyses determined that AGGTGT (-1254/-1249) is the core sequence for NF-kappaB-binding and activation. Biologically, tumor necrosis factor-alpha (TNF-alpha) activated endogenous PAcP expression in LNCaP human prostate cancer cells. The data collectively indicate that NF-kappaB up-regulates PAcP promoter activity via its binding to the AGGTGT motif, a novel binding sequence located inside the cis-active enhancer element in human prostate cancer cells.
Zelivianski
Stanislav
S
Department of Biochemistry and Molecular Biology, College of Medicine, University of Nebraska Medical Center, Omaha NE 68198, USA.
Glowacki
Richard
R
Lin
Ming-Fong
MF
D024506:5' Flanking Region; D000135:Acid Phosphatase; D001483:Base Sequence; D001665:Binding Sites; D045744:Cell Line, Tumor; D018983:DNA Footprinting; D024202:Electrophoretic Mobility Shift Assay; D004742:Enhancer Elements, Genetic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D007375:Interleukin-1 / Q000494:pharmacology; D008297:Male; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D009841:Oligonucleotides / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011467:Prostate / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000235:genetics*; D015533:Transcriptional Activation; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D020672:I-kappa B Proteins; D007375:Interleukin-1; D016328:NF-kappa B; D009841:Oligonucleotides; D014409:Tumor Necrosis Factor-alpha; D000135:Acid Phosphatase; C428439:prostatic acid phosphatase; D017027:Protein Tyrosine Phosphatases