School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, 61005, Korea.
Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania, 17033, USA.
Sci Rep. 2018 Nov 7;8(1):16481. doi: 10.1038/s41598-018-34802-z.
Swiprosin-1 is expressed in various types of cells or tissues of different species. To investigate the mechanisms underlying Swiprosin-1 expression pattern, we analyzed the promoter activity of 2-kilobase genomic sequences located at 5' flanking region of the Swiprosin-1 gene. The -2000/+41 bp of 5' flanking untranslated promoter region of Swiprosin-1 gene was constitutively transactivated without significant effect of PMA, A23187, or PMA/A23187 stimulation in Jurkat T cells. Further, we identified 5' deletant of proximal promoter region (-100/+41 to -70/+41) plays a pivotal role in activating the Swiprosin-1 gene in Jurkat T cells. Our studies also verified that ADR1 and Sp1 transcription factors were located between -70 and -100 locus of 5' flanking proximal promoter region, which is critical for the Swiprosin-1 promoter activity. ADR1 and Sp1 were shown to bind the regions of -82, -79, -76, -73 and -70 and; -79, -78 and -77, respectively, within the proximal promoter region of Swiprosin-1. Finally conserved noncoding sequences (CNS) -1, -2 and -3 were located between the exon 1 and exon 2 of Swiprosin-1 gene and synergistically transactivated the Swiprosin-1 promoter. In summary, Swiprosin-1 was constitutively expressed in Jurkat T cells by the coordinate action of ADR1 and SP1 transcription factors at the transcriptional level and CNS further boost the proximal region of Swiprosin-1 promoter activity. Our findings provide novel insights that the transcriptional regulation of Swiprosin-1 by targeting ADR1 and Sp1 binding sites may be helpful in exploring novel therapeutic strategies for advanced immune or other disorders.
Swiprosin-1 在不同物种的各种类型细胞或组织中表达。为了研究 Swiprosin-1 表达模式的机制,我们分析了位于 Swiprosin-1 基因 5' 侧翼区的 2 千碱基基因组序列的启动子活性。Swiprosin-1 基因 5' 侧翼非翻译启动子区的-2000/+41bp 在 Jurkat T 细胞中不受 PMA、A23187 或 PMA/A23187 刺激的显著影响,构成性转录激活。此外,我们确定近端启动子区域的 5' 缺失(-100/+41 至-70/+41)在 Jurkat T 细胞中激活 Swiprosin-1 基因中起着关键作用。我们的研究还验证了 ADR1 和 Sp1 转录因子位于 5' 侧翼近端启动子区域的-70 和-100 位,这对于 Swiprosin-1 启动子活性至关重要。ADR1 和 Sp1 分别被证明与 Swiprosin-1 近端启动子区域的-82、-79、-76、-73 和-70 以及-79、-78 和-77 结合。最后,保守非编码序列(CNS)-1、-2 和-3 位于 Swiprosin-1 基因的外显子 1 和外显子 2 之间,并协同激活 Swiprosin-1 启动子。总之,ADR1 和 SP1 转录因子在转录水平上的协同作用,以及 CNS 进一步增强 Swiprosin-1 启动子的近端区域,使 Swiprosin-1 在 Jurkat T 细胞中持续表达。我们的发现提供了新的见解,即靶向 ADR1 和 Sp1 结合位点对 Swiprosin-1 的转录调控可能有助于探索针对先进免疫或其他疾病的新的治疗策略。