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Cloning and Preliminary Identification of Human ACER2 Promoter
Zhu Liu 1,2, Qing Ai1,2, Huan Lan1,2, Ying Ji1,2, Zheng-Mei Yang2 , Jiang-Yi He2 , Xiao-Lu Hao2,Fang-Zhou Song1,2, You-Quan Bu1,2*
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: Alkaline ceramidase 2， ACER2， is a ceramide-hydrolyzing enzyme which plays an important role in cell proliferation， senescence， and apoptosis etc. To further investigate its transcriptional regulatory mechanism， ACER2 gene promoter has been cloned and identified in the present study. At first， the transcriptional start sites for ACER2 gene have been identified by 5' RACE (rapid amplification of cDNA ends) and bioinformatic analysis. Furthermore， three overlapping genomic fragments from the 5'- flanking region of ACER2 gene have been coloned into pGL3-basic vector to construct ACER2 promoter reporters. Luciferase reporter assay indicated that ACER2 promoter region is mainly located in a 670 bp region nearby the transcriptional start site. Transcriptional factor binding analysis indicated that， ACER2 gene promoter contains several putative binding sites for transcriptional factors such as Sp1 and GATA-1. These results suggested that transcriptional factors such as Sp1 and GATA-1 might be invovled in the transcriptional regualtion of ACER2 gene.
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