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Molecular analysis of the prostate‐specific antigen upstream gene enhancer
Author(s) -
Farmer George,
Connolly E. Sander,
Mocco J.,
Freedman Leonard P.
Publication year - 2001
Publication title -
the prostate
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.295
H-Index - 123
eISSN - 1097-0045
pISSN - 0270-4137
DOI - 10.1002/1097-0045(200101)46:1<76::aid-pros1011>3.0.co;2-4
Subject(s) - lncap , enhancer , transfection , biology , microbiology and biotechnology , androgen receptor , prostate specific antigen , reporter gene , cell culture , gene , antigen , gene expression , cancer research , prostate cancer , genetics , cancer
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. Prostate 46:76–85, 2001. © 2001 Wiley‐Liss, Inc.