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SP1 plays a pivotal role for basal activity of TIGAR promoter in liver cancer cell lines

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Abstract

TIGAR expression resulted in down-regulation of glycolysis, reduction of intracellular levels of reactive oxygen species, and protection from apoptosis. Despite biological importance, its promoter has not yet been characterized. In this study, we characterized that transcription factor SP1 plays a pivotal role for basal activity of TIGAR promoter. By 5′RACE, the transcription start site was identified locating at 134 bp upstream of the translation initiation site. Different portions of 5′-flanking and 5′-untranslated regions were fused to a luciferase reporter gene to create reporter plasmids, and constructs were transiently transfected into HepG2, Bel-7402, and Smmc-7721 cell lines for luciferase analysis. A minimal region −56/−4 bearing a SP1-binding site was characterized and plays a vital role. Data from electrophoretic mobility shift assay and chromatin immunoprecipitation showed that SP1 can interact with the SP1-binding site within TIGAR promoter in vitro and in vivo. Conclusively, SPl is indispensable for basal activity of TIGAR promoter.

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Abbreviations

RACE:

Rapid amplification of cDNA ends

TSS:

Transcriptional start site

EMSA:

Electrophoretic mobility shift assays

ChIP:

Chromatin immunoprecipitation

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Acknowledgment

We are grateful to Prof Jingyi Hui (Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) for providing research facilities and technical assistance.

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Authors and Affiliations

  1. Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200025, People’s Republic of China

    Shubiao Zou, Zhidong Gu, Jiayi Wang & Qishi Fan

  2. Faculty of Medical Laboratory Science, Shanghai Jiaotong University School of Medicine, Shanghai, 200025, People’s Republic of China

    Peihua Ni & Xiangfan Liu

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  1. Shubiao Zou
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  2. Zhidong Gu
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  3. Peihua Ni
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  4. Xiangfan Liu
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  5. Jiayi Wang
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  6. Qishi Fan
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Correspondence to Qishi Fan.

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Zou, S., Gu, Z., Ni, P. et al. SP1 plays a pivotal role for basal activity of TIGAR promoter in liver cancer cell lines. Mol Cell Biochem 359, 17–23 (2012). https://doi.org/10.1007/s11010-011-0993-0

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  • DOI: https://doi.org/10.1007/s11010-011-0993-0

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