Characterization of the promoter of 1A6/DRIM, a novel cancer-related gene and identification of its transcriptional activator.
1A6/DRIM (Down-regulated in Metastasis) has been reported to express at a high level in the gastric cancer tissues and the premalignant lesions implicating the involvement of 1A6/DRIM in cell transformation. Although the information regarding the putative functions and distribution of the 1A6/DRIM in different tissues and cell lines has been increasing recently, its promoter and promoter-regulating factors remain unknown. In this study, the transcription initiation site of 1A6/DRIM was confirmed to be located at 147 bp upstream of the ATG codon using the primer extension analysis. The minimal promoter region of the 1A6/DRIM is located between -47 and +42 of the transcription initiation site measured by luciferase reporter assays using a set of deletion constructs. In addition, an E-box is shown to be an essential element for transcriptional regulation of 1A6/DRIM demonstrated by luciferase assay with different deletion and mutation constructs. Finally, a transcription factor, upstream stimulatory factor 2 (USF2) was found to be an activator of the 1A6/DRIM through binding to the E-box demonstrated by luciferase reporter assay, electrophoretic mobility shift assay, and chromatin immunoprecipitation (ChIP) assay. The structural analysis of the 1A6/DRIM promoter and the identification of its potential regulatory effecter may help us to understand its biological functions in regulating cancer development.
Xing
Xiaoyan
X
Beijing Institute for Cancer Research, School of Oncology, Peking University, No. 1 Da Hong Luo Chang St., West District, Beijing 100034, P.R. China.
Du
Xiaojuan
X
Lu
Zheming
Z
Ning
Tao
T
Su
Xiulan
X
Ke
Yang
Y
D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D011506:Proteins / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D012261:Ribonucleoproteins; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D024363:Transcription Initiation Site; D014162:Transfection; D051720:Upstream Stimulatory Factors
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D015534:Trans-Activators; D014157:Transcription Factors; C497184:USF2 protein, human; C113527:UTP20 protein, human; D051720:Upstream Stimulatory Factors; D008156:Luciferases