RNA聚合酶II触发环在底物选择中发挥作用，并直接受到α-鹅膏蕈碱的靶向作用。
The RNA polymerase II trigger loop functions in substrate selection and is directly targeted by alpha-amanitin.
作者信息
Kaplan Craig D, Larsson Karl-Magnus, Kornberg Roger D
机构信息
Department of Structural Biology, Stanford University, Stanford, CA 94305, USA.
出版信息
Mol Cell. 2008 Jun 6;30(5):547-56. doi: 10.1016/j.molcel.2008.04.023.
Abstract
Structural, biochemical, and genetic studies have led to proposals that a mobile element of multisubunit RNA polymerases, the Trigger Loop (TL), plays a critical role in catalysis and can be targeted by antibiotic inhibitors. Here we present evidence that the Saccharomyces cerevisiae RNA Polymerase II (Pol II) TL participates in substrate selection. Amino acid substitutions within the Pol II TL preferentially alter substrate usage and enzyme fidelity, as does inhibition of transcription by alpha-amanitin. Finally, substitution of His1085 in the TL specifically renders Pol II highly resistant to alpha-amanitin, indicating a functional interaction between His1085 and alpha-amanitin that is supported by rerefinement of an alpha-amanitin-Pol II crystal structure. We propose that alpha-amanitin-inhibited Pol II elongation, which is slow and exhibits reduced substrate selectivity, results from direct alpha-amanitin interference with the TL.
摘要
结构、生化及遗传学研究表明，多亚基RNA聚合酶的一个可移动元件——触发环（TL），在催化过程中起关键作用，且可被抗生素抑制剂靶向作用。在此，我们提供证据表明，酿酒酵母RNA聚合酶II（Pol II）的TL参与底物选择。Pol II的TL内的氨基酸替换优先改变底物使用情况和酶保真度，α-鹅膏蕈碱抑制转录时也会如此。最后，TL中His1085的替换使Pol II对α-鹅膏蕈碱具有高度抗性，这表明His1085与α-鹅膏蕈碱之间存在功能相互作用，α-鹅膏蕈碱-Pol II晶体结构的重新精修也支持这一点。我们认为，α-鹅膏蕈碱抑制的Pol II延伸缓慢且底物选择性降低，是由于α-鹅膏蕈碱直接干扰了TL。
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