BmTBP upregulates the transcription of BmSuc1 in silkworm (Bombyx mori) by binding to BmTfΙΙA-S
BmTBP upregulates the transcription of BmSuc1 in silkworm (Bombyx mori) by binding to BmTfΙΙA-S
复制标题
BmTBP 通过与 BmTfÎÎA-S 结合上调家蚕 (Bombyx mori) 中 BmSuc1 的转录
DOI：
10.1111/1744-7917.13168
复制
发表时间：
2022
期刊：
影响因子：
4
通讯作者：
Meng Y
中科院分区：
文献类型：
--
作者：
Zhang XW;Yang LL;Gan Q;Jiang S;Liang D;Gao JS;Meng Y
BmSuc1基因编码一种新的动物型β-呋喃果糖苷酶(EC 3.2.1.26)，首次在家蚕中被克隆和鉴定。作为一种必需的蔗糖酶，BmSUC1的活性不受桑叶中生物碱类糖类物质的影响。这种酶还可以直接调节家蚕中肠中蔗糖的水解度。此外，BmSUC1还参与丝腺组织中丝胶1的合成。然而，BmSuc1转录调控的机制尚不清楚。在这项研究中，我们使用双荧光素酶报告基因分析了BmSuc1启动子的活性，并确定了4个对转录激活至关重要的区域。克隆了编码能与核心启动子区域结合的转录因子(TATA盒结合蛋白；BmTBP)的基因。实时定量聚合酶链式反应分析表明，该基因在中肠高表达。通过RNA干扰下调BmTbP的表达，可降低BmSuc1在转录和蛋白水平的表达。电泳迁移率改变分析和染色质免疫沉淀表明，BmTBP能与BmSuc1启动子中的TATA盒顺式调节元件结合。此外，基于生物信息学的分析和远距离蛋白质印迹揭示了BmTBP与另一转录因子(BmTfIIA-S)之间的相互作用。荧光素酶报告基因分析结果证实，BmTBP-BmTfIIA-S复合体提高了BmSuc1启动子的活性。综上所述，这些发现表明，BmTBP通过与BmTfIIA-S相互作用来调节BmSuc1的表达。
TheBmSuc1gene, which encodes a novel animal‐typeβ‐fructofuranosidase (EC 3.2.1.26), was first cloned and identified in silkworm (Bombyx mori). As an essential sucrase, the activity of BmSUC1 is unaffected by alkaloidal sugar mimics in mulberry leaves. This enzyme may also directly regulate the degree of sucrose hydrolysis in the silkworm midgut. In addition, BmSUC1 is involved in the synthesis of sericin 1 in the silk gland tissue. However, the mechanism underlying the regulation ofBmSuc1transcription remains unclear. In this study, we analyzed theBmSuc1promoter activity using a dual‐luciferase reporter assay and identified 4 regions that are critical for transcriptional activation. The gene encoding a predicted transcription factor (TATA‐box‐binding protein; BmTBP) capable of binding to the core promoter regions was cloned. A quantitative real‐time polymerase chain reaction analysis indicated the gene was highly expressed in the midgut. DownregulatingBmTBPexpression via RNA interference decreased the expression ofBmSuc1at the transcript and protein levels. An electrophoretic mobility shift analysis and chromatin immunoprecipitation indicated that BmTBP can bind to the TATA‐boxcis‐regulatory element in theBmSuc1promoter. Furthermore, a bioinformatics‐based analysis and a far‐western blot revealed the interaction between BmTBP and another transcription factor (BmTfIIA‐S). The luciferase reporter gene assay results confirmed that the BmTBP–BmTfIIA‐S complex increases theBmSuc1promoter activity. Considered together, these findings suggest that BmTBP regulatesBmSuc1expression through its interaction with BmTfIIA‐S.