title: Molecular analysis of the prostate-specific antigen upstream gene enhancer.
abstract: BACKGROUND
  Our objective was to identify factors other than androgen receptor that bind to and regulate the prostate-specific antigen (PSA) upstream gene enhancer (PSE).
  
  
  METHODS
  DNAse I footprinting and electromobility shift assays (EMSA) were performed over the PSE using lysates from PSA-producing cell lines, LNCaP and LAPC4, and nonproducing PSA cell lines, PC-3 cells, U937 monocytes, and Namalwa B cells. Mutational analysis and transient transfection assays were used to determine the contributions made by different elements towards the regulation of the enhancer.
  
  
  RESULTS
  Three distinct regions surrounding androgen response elements of the PSE were found to bind unknown ubiquitous and cell type-specific proteins. These regions, when mutated in a PSE reporter construct, were shown to be required for maximal activation in LNCaP cells.
  
  
  CONCLUSIONS
  These results correlate unknown sequence-specific DNA binding proteins with androgen-mediated regulation of a prostate-specific gene, thus providing further insight into the mechanism of PSA gene expression.
authors.affiliation: Cell Biology Program, Memorial Sloan-Kettering Cancer Institute, New York, NY, USA.
authors.forename: G
authors.initials: G
authors.lastname: Farmer
authors.forename: E S
authors.initials: ES
authors.lastname: Connolly
authors.forename: J
authors.initials: J
authors.lastname: Mocco
authors.forename: L P
authors.initials: LP
authors.lastname: Freedman
mesh_terms: D001483:Base Sequence; D018983:DNA Footprinting; D017931:DNA Primers / Q000737:chemistry; D003850:Deoxyribonuclease I / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000737:chemistry; D017430:Prostate-Specific Antigen / Q000737:chemistry / Q000235:genetics*; D011471:Prostatic Neoplasms / Q000737:chemistry / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D017931:DNA Primers; D003850:Deoxyribonuclease I; D017430:Prostate-Specific Antigen