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粗糙脉孢菌核糖体蛋白基因和核糖体RNA基因共有的功能性启动子元件。

Functional promoter elements common to ribosomal protein and ribosomal RNA genes in Neurospora crassa.

作者信息

Cujec T P, Tyler B M

机构信息

Department of Plant Pathology, University of California, Davis 95616, USA.

出版信息

Mol Gen Genet. 1996 Nov 27;253(1-2):205-16. doi: 10.1007/s004380050314.

Abstract

The promoter sequences of a cytoplasmic ribosomal protein gene (crp-2) of Neurospora crassa were identified using promoter deletion and substitution mutants. A gene-targeting strategy was used to assay the mutants in vivo. The promoter architecture of crp-2 is complex and is more similar to that of ribosomal protein genes in mouse than in Saccharomyces cerevisiae. Six regions were identified as important for transcription. These included two elements, a CG repeat and a Dde box, that are conserved in most other promoters of N. crassa ribosomal protein genes and have also been demonstrated as being required for transcription from the 40 S rRNA promoter by RNA polymerase I in vitro. The CG repeats located at -73 to -66 and between -189 and -154 were functionally redundant and increased transcription efficiency by 10- to 15- fold. The Dde boxes located at -153 to -147 and at -95 to -83 contributed 2-fold and 5-fold to transcription efficiency, respectively. An unidentified element between -254 and -190 contributed 2-fold, while a pyrimidine-rich region between -85 and -66 influenced the start point of transcription.

摘要

利用启动子缺失和替代突变体鉴定了粗糙脉孢菌细胞质核糖体蛋白基因(crp - 2)的启动子序列。采用基因靶向策略在体内分析这些突变体。crp - 2的启动子结构复杂,与小鼠核糖体蛋白基因的启动子结构相比,比酿酒酵母的更相似。确定了六个对转录重要的区域。其中包括两个元件,一个CG重复序列和一个Dde框,它们在粗糙脉孢菌核糖体蛋白基因的大多数其他启动子中保守,并且在体外也已证明是RNA聚合酶I从40 S rRNA启动子转录所必需的。位于 - 73至 - 66以及 - 189至 - 154之间的CG重复序列在功能上是冗余的,可将转录效率提高10至15倍。位于 - 153至 - 147以及 - 95至 - 83的Dde框分别对转录效率贡献了2倍和5倍。位于 - 254至 - 190之间的一个未鉴定元件贡献了2倍,而位于 - 85至 - 66之间的富含嘧啶的区域影响转录起始点。

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