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Identification and characterization of human PCDH10 gene promoter

Author:
Zesong Lia   Jun Xiea   Wenjie Lia   Aifa Tanga   Xianxin Lia   Zhimao Jianga   Yonghua Hana   Jiongxian Yea   Jie Jingb   Yaoting Guia   guiyaoting2007@yahoo.com.cn  Zhiming Caia   c   caizhiming2000@yahoo.com.cn  


Journal:
Gene


Issue Date:
2011


Abstract(summary):

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.


Page:
49-56


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