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Preliminary Identification and Characterization of Human Ska2 Core Promoter Region
Ai Qing1,2, Weng Huali1,2, Zhang Ying1,2, Liu Zhu1,2, Wang Yitao1,2, Li Yi1,2,Zhu Huifang1,2, Zhang Chundong1,2*, 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: Ska2 (spindle and KT associated 2)， also known as FAM33A (family with sequence similarity 33， member A)， is a recently discovered novel gene involved in both cell cycle regulation and tumorigenesis. The molecular regulatory mechanism of its expression， however， remains unclear. We have previously identified its main promoter region， thus the present study was designed to further identify and characterize its core promoter region. PCR was used to make five different deletion mutants of Ska2 promoter reporter. Either or both of the two potential NF-Y binding sites were mutated to make three Ska2 mutant promoter reporters using PCR based sitedirected mutagenesis. Dual luciferase reporter assay was used to determine promoter reporter activity. The results of promoter activity analysis showed that the five deletion mutants showed significant promoter activity， and sitedirected mutagenesis of either or both NF-Y binding sites resulted in decreased promoter activity. Additionally，ectopic overexpression of NF-Y enhanced the Ska2 promoter activity. Taken together， our results suggested that the core promoter region of Ska2 gene is mainly located in a 80 bp region and NF-Y might play an important role in the transcriptional regulation of Ska2 gene.
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