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由Rtg1p介导的反式激活,Rtg1p是一种碱性螺旋-环-螺旋蛋白,在酵母线粒体与细胞核之间的通讯中发挥作用。

Transactivation by Rtg1p, a basic helix-loop-helix protein that functions in communication between mitochondria and the nucleus in yeast.

作者信息

Rothermel B A, Shyjan A W, Etheredge J L, Butow R A

机构信息

Department of Biochemistry, University of Texas, Southwestern Medical Center, Dallas 75235, USA.

出版信息

J Biol Chem. 1995 Dec 8;270(49):29476-82. doi: 10.1074/jbc.270.49.29476.

Abstract

Rtg1p is a basic helix-loop-helix transcription factor in the yeast Saccharomyces cerevisiae that is required for basal and regulated expression of CIT2, the gene encoding a peroxisomal isoform of citrate synthase. In respiratory incompetent rho degree petite cells, CIT2 transcription is elevated as much as 30-fold compared with respiratory competent rho + cells. Here we provide evidence that Rtg1p interacts directly with a CIT2 upstream activation site (UASr) and that the rho degree/rho + regulation is not due to a change in the levels of Rtg1p. A fusion protein consisting of the DNA binding domain of Gal4p fused to the NH2 terminus of the full-length wild-type Rtg1p was able to transactivate an integrated LacZ reporter under control of the Gal4p-responsive GAL1 UASG in a rho degree/rho(+)-dependent manner. Other Gal4p fusions to deletions or mutations of Rtg1p indicate that the helix-loop-helix domain is essential for transactivation. Regulated expression of CIT2 also requires the RTG2 gene product. The Gal4-Rtg1p fusion was unable to transactivate the LacZ reporter gene in a strain deleted for RTG2, suggesting that the RTG2 product does not act independently of Rtg1p in the rho degree/rho + transcriptional response.

摘要

Rtg1p是酿酒酵母中的一种碱性螺旋-环-螺旋转录因子,它是CIT2基因基础表达和调控表达所必需的,CIT2基因编码柠檬酸合酶的一种过氧化物酶体同工型。在呼吸功能不全的ρ⁰小菌落细胞中,与呼吸功能正常的ρ⁺细胞相比,CIT2转录水平升高多达30倍。在这里,我们提供证据表明Rtg1p直接与CIT2上游激活位点(UASr)相互作用,并且ρ⁰/ρ⁺调控并非由于Rtg1p水平的变化。一种由Gal4p的DNA结合结构域与全长野生型Rtg1p的NH2末端融合而成的融合蛋白,能够以ρ⁰/ρ⁺依赖性方式在Gal4p反应性GAL1 UASG的控制下反式激活整合的LacZ报告基因。其他与Rtg1p缺失或突变的Gal4p融合表明,螺旋-环-螺旋结构域对于反式激活至关重要。CIT2的调控表达也需要RTG2基因产物。Gal4-Rtg1p融合蛋白在缺失RTG2的菌株中无法反式激活LacZ报告基因,这表明RTG2产物在ρ⁰/ρ⁺转录反应中并非独立于Rtg1p发挥作用。

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