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Published online 30 April 2004

Nucleic Acids Research, 2004, Vol. 32, No. 8 2482-2493
© 2004 Oxford University Press

A DNA replication-related element downstream from the initiation site of Drosophila selenophosphate synthetase 2 gene is essential for its transcription

Jing Shun Jin, Seunghee Baek, Hyesin Lee, Mi Young Oh, Yong Eui Koo, Myoung Sup Shim, So Yeon Kwon, Iksoo Jeon1, So Young Park2, Kwanghee Baek1, Mi Ae Yoo2, Dolph Lee Hatfield3 and Byeong Jae Lee*

School of Biological Sciences and Institute of Molecular Biology and Genetics, Seoul National University, Korea, 1 Graduate School of Biotechnology, Kyung Hee University, Yongin, Korea, 2 Department of Molecular Biology, Pusan National University, Busan, Korea and 3 Molecular Biology of Selenium Section, Laboratory of Cancer Prevention, Center for Cancer Research, National Cancer Institute, National Institutes of Health, MD, USA

*To whom correspondence should be addressed. Tel: +82 2 880 6775; Fax: +82 2 872 9019; Email: imbglmg{at}plaza.snu.ac.kr

Received January 25, 2004; Revised March 27, 2004; Accepted April 6, 2004

Selenophosphate synthetase catalyzes the synthesis of selenophosphate which is a selenium donor for Sec biosynthesis. In Drosophila melanogaster, there are two types of selenophosphate synthetases designated dSPS1 and dSPS2, where dSPS2 is a selenoprotein. The mechanism of gene expression of dSPS2 as well as other selenoproteins in Drosophila has not been elucidated. Herein, we report an essential regulator system that regulates the transcription of the dSPS2 gene (dsps2). Through deletion/substitution mutagenesis, the downstream DNA replication-related element (DRE) located at +71 has been identified as an essential element for dsps2 promoter activity. Furthermore, double-stranded RNA interference (dsRNAi) experiments were performed to ablate transcription factors such as TBP, TRF1, TRF2 and DREF in Schneider cells. The dsRNAi experiments showed that dsps2 promoter activities in DREF- and TRF2-depleted cells were significantly decreased by 90% and 50%, respectively. However, the depletion of TBP or TRF1 did not affect the expression level of dsps2 even though there is a putative TATA box at –20. These results strongly suggest that the DRE/DREF system controls the basal level of transcription of dsps2 by interacting with TRF2.


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