Regulation of SLITRK1 gene by neuron restrictive silencer factor in NMB cells
김명미, 김진수, 문성민, 최미숙, 박보람, 이동설, 모신엽, 조선호, 김춘성,
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김명미 ( Kim Myung-Mi ) - 조선대학교 치의학전문대학원 구강생화학교실
김진수 ( Kim Jin-Soo ) - 조선대학교 치의학전문대학원 구강생물학연구소
문성민 ( Moon Sung-Min ) - 조선대학교 치의학전문대학원 구강생화학교실
최미숙 ( Choi Mi-Suk ) - 조선대학교 치의학전문대학원 구강생화학교실
박보람 ( Park Bo-Ram ) - 조선대학교 치의학전문대학원 구강생화학교실
이동설 ( Lee Dong-Seol ) - 조선대학교 치의학전문대학원 구강생물학연구소
모신엽 ( Mo Shin-Yeob ) - 조선대학교 치의학전문대학원 구강생물학연구소
조선호 ( Cho Seon-Ho ) - 조선대학교 치의학전문대학원 구강생물학연구소
김춘성 ( Kim Chun-Sung ) - 조선대학교 치의학전문대학원 구강생물학연구소
KMID : 0353320130370020088
Abstract
Purpose: The Slitrk family are neuronal transmembrane proteins that participate in the regulation of neurite outgrowth and are selectively expressed in the brains of adult mice. Neuron restrictive silencer factor (NRSF), also called RE1-silencing transcription factor, was originally discovered as a transcriptional repressor in non-neuronal cells. However, in this study, we show that the neuron restrictive silencer element (NRSE) of SLITRK1 functions as a critical regulator to repress SLITRK1 gene expression in NMB cells.
Materials and Methods: To screen NRSF/NRSE-regulated specific genes, we performed microarray analysis of NRSF siRNA-transfected NMB cells. We selected eight genes and confirmed their expression by RT-PCR analysis.
Results: Expression levels of SLITRK1, VGF, and CCNB1 were up-regulated in NRSF siRNA-transfected NMB cells compared with control NMB cells, whereas TGFBR3, CDC25B, E2F1, TGFB1, and TNFRSF21 were slightly down-regulated or showed no change. Especially, we identified three putative NRSF-binding sequences in the SLITRK1 gene, and its transcription was significantly up-regulated in NRSF siRNA-transfected NMB cells as measured by real-time PCR. Using co-transfection studies, we determined that the second NRSE of the SLITRK1 gene was functional in NMB cells. In addition, NRSF bound to the second NRSE of the SLITRK1 gene in a sequence-specific manner as confirmed by supershift assay. Our data suggest that NRSF interacts with and represses expression of the SLITRK1 gene.
Conclusion: NRSF plays an important role in regulating the growth and development of neuronal genes via SLITRK1 gene transcription.
키워드
RE1-silencing transcription factor; Neuron restrictive silencer element; Transcriptional repressor; SLITRK1
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