Abstract
The hepatocellular carcinoma suppressor 1 (HCCS1) gene was identified by both positional cloning from a predominant region of loss of heterozygosity (17p13.3) in liver cancer and by functional screening for genes affecting cell proliferation in large-scale transfection assays. Its overexpression results in inhibition of cell proliferation in cell culture and tumor growth in nude mice. To understand its transcription regulation, the promoter architecture has been dissected in detail. The major start of transcription was mapped by primer extension to a C residue, 177 nucleotides upstream of the ATG codon. By assessing the promoter activity of a set of linker-scanning mutants of the minimal promoter (−60 to +148 region) in a transient transfection assay, we found that the +1 to + 40 region is critical to HCCS1 gene transcription, containing binding sites for transcription factors NF-κB (−21 to +7 and +40 to +26), p53 (+29 to +9) and ETS (+4 to +20 and +23 to +39). Biochemical and molecular analyses revealed that the ETS transcription factors ETS-2 and Elf-1 bind to the two ETS sites in situ and contribute significantly to the transcriptionally active state of the HCCS1 gene, while NF-κB, p53 and two other members of the ETS family (ETS-1 and NERF2) appear to play little role. Our observations provide insight into the mechanistic aspects of HCCS1 transcription regulation.
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Acknowledgements
Thanks are due to Y Shi and W Heniig for their comments on this manuscript, as well as H Ji and N Rice for their kind provision of constructs. This work was supported by grants to Jing De Zhu from the Shanghai Science Foundation (04DZ14006 and 05DZ19318), the National Science Foundation (30450001, 30570850 and 10574134) and the National Research Program for Basic Research of China (2004CB518804 and 2002CB713700).
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Zhu, J., Fei, Q., Wang, P. et al. Transcription of the putative tumor suppressor gene HCCS1 requires binding of ETS-2 to its consensus near the transcription start site. Cell Res 16, 780–796 (2006). https://doi.org/10.1038/sj.cr.7310092
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DOI: https://doi.org/10.1038/sj.cr.7310092
