title: Identification and characterization of human PCDH10 gene promoter.
abstract: Recent studies have suggested roles for PCDH10 as a novel tumor suppressor gene. In our previous work, we located the core promoter of PCDH10 to a 462-bp segment of 5'-flanking region characterized by a high GC content. Here we further identified and characterized the promoter for PCDH10. Transient transfection of PC3 and LNCaP cells with a series of deleted promoter constructs indicated that the minimal promoter region was between nucleotides -144 and -99. This segment contained a CAAT box, a GT box, and a putative transcription factor binding site for AP-4. Mutational analysis identified that the CAAT box and GT box are necessary for promoter activity. Ectopic expression of NF-Ys increased reporter gene activity, whereas expression of a dominant-negative NF-YA decreased reporter gene activity. Co-transfection of Sp1/Sp3 expression plasmids enhanced reporter gene activity in a dose-dependent manner. Mithramycin A, an inhibitor of Sp-DNA interaction, reduced PCDH10 promoter activity. Electrophoretic mobility shift assays and chromatin immunoprecipitation demonstrated binding of transcription factors Sp1/Sp3 to the promoter region in vitro and in vivo. Our data show that Sp1/Sp3 and CBF/NF-Y transcription factors play a crucial role in the basal expression of the human PCDH10 gene.
authors.affiliation: Guangdong Key Laboratory of Male Reproductive Medicine and Genetics, Peking University Shenzhen Hospital, Shenzhen PKU-HKUST Medical Center, 1120 Lianhua Road, Shenzhen, Guangdong, PR China.
authors.forename: Zesong
authors.initials: Z
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Xie
authors.forename: Wenjie
authors.initials: W
authors.lastname: Li
authors.forename: Aifa
authors.initials: A
authors.lastname: Tang
authors.forename: Xianxin
authors.initials: X
authors.lastname: Li
authors.forename: Zhimao
authors.initials: Z
authors.lastname: Jiang
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Han
authors.forename: Jiongxian
authors.initials: J
authors.lastname: Ye
authors.forename: Jie
authors.initials: J
authors.lastname: Jing
authors.forename: Yaoting
authors.initials: Y
authors.lastname: Gui
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Cai
mesh_terms: D001483:Base Sequence; D023081:CCAAT-Binding Factor / Q000378:metabolism; D015820:Cadherins / Q000235:genetics*; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D004252:DNA Mutational Analysis; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D000091363:Protocadherins; D016329:Sp1 Transcription Factor / Q000378:metabolism; D051705:Sp3 Transcription Factor / Q000378:metabolism
chemical_list: D023081:CCAAT-Binding Factor; D015820:Cadherins; C492061:NFYA protein, human; C438603:PCDH10 protein, human; D000091363:Protocadherins; D016329:Sp1 Transcription Factor; D051705:Sp3 Transcription Factor