Exp Ther Med. 2013 Nov;6(5):1290-1294. doi: 10.3892/etm.2013.1275. Epub 2013 Aug 28.

Analysis of promoters and CREB/AP-1 binding sites of the human TMEM174 gene.

Experimental and therapeutic medicine

Fen Hu, Yanan Meng, Lixia Gou, Xiujun Zhang

PMID: 24223660 PMCID: PMC3820833 DOI: 10.3892/etm.2013.1275

Abstract

Transmembrane protein 174 (TMEM174) is a type III transmembrane protein with no clear signal peptide. The N and C terminals are located inside the cell. Our previous study demonstrated high expression of TMEM174 in the kidney and its potential involvement in renal cancer based on its capacity to stimulate cell proliferation. However, the mechanism by which TMEM174 promotes proliferation at the transcriptional level remains to be elucidated. In the present study, the TMEM174 promoter region was amplified from whole blood DNA. Six different regions of the regulatory sequences of the TMEM174 promoter region including ~2.5 kb of the upstream region were cloned into the dual luciferase expression vector pGL3-basic. Comparison of the activity of these fragments in dual luciferase reporter assays revealed higher levels of activity for the fragments spanning -186 to +674, -700 to +674, -1,000 to +674 and -2,500 to +1 bp. Lower levels of activity were detected for the fragments spanning -466 to +674 and -890 to +674 bp. The highest activity was detected for the fragment spanning -186 to +674 bp. Electrophoretic mobility shift assay (EMSA) was performed to determine effective transcription factor binding sites. Specific binding of the cyclic-AMP response element binding (CREB) within the TMEM174 gene promoter region was demonstrated, although binding of the activator protein-1 (AP-1) was also detected in this region. In conclusion, these results suggest that the core promoter region of the human TMEM174 gene is located within the region spanning -186 to +674 bp and that the transcription factors CREB and AP-1 are involved in the transcriptional regulation of this gene.

Keywords: activator protein-1; core promoter; cyclic-AMP response element binding protein; luciferase; transcriptional regulation; transmembrane protein 174 gene

References

1. Antioxid Redox Signal. 2005 Nov-Dec;7(11-12):1648-63 - PubMed
2. Endocr Rev. 2001 Apr;22(2):153-83 - PubMed
3. Cancer Res. 2012 Mar 15;72(6):1449-58 - PubMed
4. Future Oncol. 2007 Aug;3(4):475-80 - PubMed
5. Biochem Biophys Res Commun. 2010 Apr 16;394(4):993-9 - PubMed
6. Oncogene. 2001 Apr 30;20(19):2390-400 - PubMed
7. Cancer Lett. 2012 Jul 28;320(2):158-70 - PubMed
8. Leuk Lymphoma. 2004 Feb;45(2):265-70 - PubMed
9. Crit Rev Oncog. 2011;16(1-2):37-46 - PubMed
10. Cell Cycle. 2005 Sep;4(9):1134-5 - PubMed
11. Food Chem Toxicol. 2012 Mar;50(3-4):558-66 - PubMed
12. Proc Natl Acad Sci U S A. 2004 Nov 2;101(44):15724-9 - PubMed
13. Nat Rev Cancer. 2003 Nov;3(11):859-68 - PubMed
14. Bioessays. 2012 Apr;34(4):293-300 - PubMed
15. PLoS One. 2012;7(2):e31193 - PubMed
16. J Biol Chem. 2002 Sep 27;277(39):35961-8 - PubMed
17. Mol Cell. 2000 Nov;6(5):1109-19 - PubMed
18. Cancer. 2005 Nov 1;104(9):1819-24 - PubMed
19. Dev Cell. 2003 Jun;4(6):865-77 - PubMed
20. Gene. 2012 Mar 15;496(1):8-16 - PubMed
21. Dev Biol. 2007 Jul 1;307(1):127-41 - PubMed

Publication Types

• Journal Article