Transcriptional Regulation of Cuticular Protein Glycine-Rich13 Gene Expression in Wing Disc of Bombyx mori, Lepidoptera
Transcriptional Regulation of Cuticular Protein Glycine-Rich13 Gene Expression in Wing Disc of Bombyx mori, Lepidoptera
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DOI：
10.1093/jisesa/iev019
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发表时间：
2015-04-05
影响因子：
2.2
通讯作者：
Swapon, Ahsanul Haque
中科院分区：
文献类型：
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作者：
Ali, Md. Saheb;Rahman, R. F.;Swapon, Ahsanul Haque
昆虫表皮蛋白基因是研究蜕皮激素信号转导分子机制的理想模型。本研究对家蚕表皮蛋白富含甘氨酸13（CPG 13）基因表达的激素调控及重要调控序列和转录因子进行了分析。CPG 13在B的翅组织中表达较强，在蛹前期表达较强。森。CPG 13的表达被20 E诱导，而在翅盘中被放线菌酮抑制。CPG 13基因的上游区域进行了分析，使用一个基因枪系统的瞬时报告分析，并确定了两个BR-Z2结合位点是重要的顺式作用元件的蜕皮激素脉冲的荧光素酶报告基因的转录激活。这些位点的定点诱变，然后引入翼盘，显着降低了报告活性。结果发现，携带蜕皮激素响应转录因子BR-Z2的结合位点的区域负责翅盘中报告基因活性的激素增强。BR-Z2结合位点的突变降低了报告活性，表明BR-Z2亚型可以结合到角质层蛋白基因CPG 13的上游区域并激活其表达。
Cuticular protein genes are good models to study the molecular mechanisms of signaling by ecdysteroids, which regulate molting and metamorphosis in insects. The present research demonstrates on hormonal regulation and analysis of the regulatory sequences and transcription factors important for Bombyx mori cuticular protein glycine-rich13 (CPG13) gene expression. Expression of CPG13 was strong at prepupal stage in wing tissues of B. mori. CPG13 expression was induced by the addition of 20E, which was inhibited by cycloheximide in the wing disc. The upstream region of the CPG13 gene was analyzed using a transient reporter assay with a gene gun system and identified two BR-Z2 binding sites to be important cis-acting elements for the transcription activation of the luciferase reporter gene by an ecdysone pulse. Site-directed mutagenesis of these sites, followed by introduction into wing discs, significantly decreased the reporter activity. It was found that the regions carrying the binding sites for the ecdysone-responsive transcription factor BR-Z2 were responsible for the hormonal enhancement of the reporter gene activity in wing discs. Mutation of the BR-Z2 binding sites decreased the reporter activity suggesting that the BR-Z2 isoform can bind to the upstream region of the cuticle protein gene, CPG13 and activates its expression.