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The Novel Transcription Factor e(y)2 Interacts with TAFII40 and Potentiates Transcription Activation on Chromatin Templates

机译:新型转录因子e(y)2与TAFII40相互作用并增强了染色质模板上的转录激活

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Weak hypomorph mutations in the enhancer of yellowgenes, e(y)1 and e(y)2, of Drosophila melanogaster were discovered during the search for genes involved in the organization of interaction between enhancers and promoters. Previously, the e(y)1 gene was cloned and found to encode TAFII40 protein. Here we cloned the e(y)2 gene and demonstrated that it encoded a new ubiquitous evolutionarily conserved transcription factor. The e(y)2 gene is located at 10C3 (36.67) region and is expressed at all stages ofDrosophila development. It encodes a 101-amino-acid protein, e(y)2. Vertebrates, insects, protozoa, and plants have proteins which demonstrate a high degree of homology to e(y)2. The e(y)2 protein is localized exclusively to the nuclei and is associated with numerous sites along the entire length of the salivary gland polytene chromosomes. Both genetic and biochemical experiments demonstrate an interaction between e(y)2 and TAFII40, while immunoprecipitation studies demonstrate that the major complex, including both proteins, appears to be distinct from TFIID. Furthermore, we provide genetic evidence suggesting that the carboxy terminus of dTAFII40 is important for mediating this interaction. Finally, using an in vitro transcription system, we demonstrate that recombinant e(y)2 is able to enhance transactivation by GAL4-VP16 on chromatin but not on naked DNA templates, suggesting that this novel protein is involved in the regulation of transcription.
机译:果蝇的黄色基因增强子 e(y)1 e(y)2 的弱亚型突变/ em>是在寻找与增强子和启动子之间相互作用的组织有关的基因时发现的。以前,克隆了 e(y)1 基因并发现其编码TAF II 40蛋白。在这里,我们克隆了 e(y)2 基因,并证明它编码了一个新的无处不在的进化保守转录因子。 e(y)2 基因位于10C3(36.67)区域,并在果蝇发育的所有阶段表达。它编码101个氨基酸的蛋白质e(y)2。脊椎动物,昆虫,原生动物和植物的蛋白质均显示与e(y)2高度同源。 e(y)2蛋白仅位于细胞核内,并与唾液腺多烯染色体的整个长度上的许多位点相关。遗传和生化实验均表明e(y)2与TAF II 40之间存在相互作用,而免疫沉淀研究表明,主要复合物(包括这两种蛋白质)似乎与TFIID不同。此外,我们提供的遗传证据表明,dTAF II 40的羧基末端对于介导这种相互作用很重要。最后,使用体外转录系统,我们证明重组e(y)2能够通过染色质而不是裸DNA模板上的GAL4-VP16增强反式激活,这表明这种新型蛋白质参与了转录的调控。

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