Rna14-Rna15组装介导酿酒酵母切割因子IA的RNA结合能力。
Rna14-Rna15 assembly mediates the RNA-binding capability of Saccharomyces cerevisiae cleavage factor IA.
作者信息
Noble Christian G, Walker Philip A, Calder Lesley J, Taylor Ian A
机构信息
Division of Protein Structur, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
出版信息
Nucleic Acids Res. 2004 Jun 23;32(11):3364-75. doi: 10.1093/nar/gkh664. Print 2004.
Abstract
The Rna14-Rna15 complex is a core component of the cleavage factor IA RNA-processing complex from Saccharomyces cerevisiae. To understand the assembly and RNA-binding properties, we have isolated and characterized the Rna14-Rna15 complex using a combination of biochemical and biophysical methods. Analysis of the purified complex, using transmission electron microscopy, reveals that the two proteins assemble into a kinked rod-shaped structure and that these rods are able to further self-associate. Analytical ultracentrifugation reveals that Rna14 mediates this association and facilitates assembly of an A2B2 tetramer (M(r) 230 000), where relatively compact Rna14-Rna15 heterodimers are in rapid equilibrium with tetramers that have a more extended shape. The Rna14-Rna15 complex, unlike the individual components, binds to an RNA oligonucleotide derived from the 3'-untranslated region of the S.cerevisiae GAL7 gene. Based on these structural and thermodynamic data, we propose that CFIA assembly regulates RNA-binding activity.
摘要
Rna14 - Rna15复合物是酿酒酵母中切割因子IA RNA加工复合物的核心组分。为了解其组装和RNA结合特性，我们结合生化和生物物理方法分离并表征了Rna14 - Rna15复合物。使用透射电子显微镜对纯化后的复合物进行分析，结果显示这两种蛋白质组装成一种扭结的棒状结构，并且这些棒状物能够进一步自我缔合。分析超速离心表明，Rna14介导这种缔合并促进A2B2四聚体（相对分子质量230 000）的组装，其中相对紧密的Rna14 - Rna15异源二聚体与形状更伸展的四聚体处于快速平衡状态。与单个组分不同，Rna14 - Rna15复合物能结合来源于酿酒酵母GAL7基因3'非翻译区的RNA寡核苷酸。基于这些结构和热力学数据，我们提出切割因子IA的组装调节RNA结合活性。
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