A DNA replication-related element downstream from the initiation site of Drosophila selenophosphate synthetase 2 gene is essential for its transcription.
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.
Jin
Jing Shun
JS
School of Biological Sciences and Institute of Molecular Biology and Genetics, Seoul National University, Korea.
Baek
Seunghee
S
Lee
Hyesin
H
Oh
Mi Young
MY
Koo
Yong Eui
YE
Shim
Myoung Sup
MS
Kwon
So Yeon
SY
Jeon
Iksoo
I
Park
So Young
SY
Baek
Kwanghee
K
Yoo
Mi Ae
MA
Hatfield
Dolph Lee
DL
Lee
Byeong Jae
BJ
D024506:5' Flanking Region; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004261:DNA Replication; D004330:Drosophila / Q000201:enzymology / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics*; D004790:Enzyme Induction; D017344:Genes, Insect; D008969:Molecular Sequence Data; D010770:Phosphotransferases / Q000235:genetics*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D020218:Response Elements; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology; D024363:Transcription Initiation Site; D015533:Transcriptional Activation
C099494:Dref protein, Drosophila; D029721:Drosophila Proteins; D014157:Transcription Factors; D010770:Phosphotransferases; C499550:Sps2 protein, Drosophila; C062762:selenophosphate synthetase; C109636:sps1 protein, Drosophila