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Site-directed Mutagenic Analysis of NF-Y-binding Sites in Human PRR11 Core Promoter
Weng Huali1,2, Zhang Ying1,2, Ai Qing1,2, Long Yinjiang1,2, Xie Mengyu1,2, Wang Yitao1,2,Cai Wei1,2, Zhu Huifang1,2*, Bu Youquan1,2*
1Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing 400016, China;2Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China
Abstract: Proline-rich protein 11 (PRR11)， a novel tumor-associated gene discovered by our group， plays an important role in cell cycle and carcinogenesis， etc. Based on the previous identification and analysis of PRR11 promotor， the present study focuses on determining the role of nuclear factor Y (NF-Y) binding sites in PRR11 core promoter region. Nucleotide sequence homology analysis indicated that， the human PRR11 gene promoter core sequence contains two NF-Y binding sites which are highly conservative in human， cow， rat and mouse. Co-transfection experiment revealed that ectopic over-expression of NF-Y could significantly increase the PRR11 promoter activity. However， efficient site-directed mutagenesis of each or both NF-Y-binding sites resulted in a significant de crease of PRR11 promoter activity and a remarkable attenuation of PRR11 promoter activation driven by the ectopic NF-Y over-expression. In addition， we also improved the conventional base-changing methods by combining the transcription factor binding prediction. Taken together， our present study strongly suggested that NF-Y binding sites are the important cis-acting elements in PRR11 core promoter region， and NF-Y might be involved in the cell cycle and carcinogenesis via regulating the transcription of human PRR11 gene.
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