Elastic coupling between RNA degradation and unwinding by an exoribonuclease.
Elastic coupling between RNA degradation and unwinding by an exoribonuclease.
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DOI：
10.1126/science.1216848
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发表时间：
2012-06-29
期刊：
影响因子：
--
通讯作者：
Ha T
中科院分区：
文献类型：
--
作者：
Lee G;Bratkowski MA;Ding F;Ke A;Ha T
Rrp44(Dis3)是酵母外切体复合体的关键催化亚基，可在3‘-5’方向上一次消化一个核苷酸。它的运动功能是由水解性核酸酶反应释放的能量提供动力，而不是像传统的螺旋酶那样由ATP水解酶提供能量。单分子荧光分析表明，Rrp44不是在单个碱基对步骤中解开RNA，而是积累多个单核苷酸步骤水解反应释放的能量，直到大约4个碱基对突然解开。动力学分析表明，RNA解离而不是切割或链释放决定了总的RNA降解率，解离步长由Rrp44/RNA复合体的非线性弹性决定，而不是由双链稳定性决定。
Rrp44 (Dis3) is a key catalytic subunit of the yeast exosome complex and can processively digest structured RNA one nucleotide at a time in the 3’ to 5’ direction. Its motor function is powered by the energy released from the hydrolytic nuclease reaction instead of ATP hydrolysis as in conventional helicases. Single-molecule fluorescence analysis revealed that instead of unwinding RNA in single base pair steps, Rrp44 accumulates the energy released by multiple single nucleotide step hydrolysis-reactions until about four base pairs are unwound in a burst. Kinetic analyses showed that RNA unwinding, not cleavage or strand release, determines the overall RNA degradation rate, and that the unwinding step size is determined by the nonlinear elasticity of the Rrp44/RNA complex, but not by duplex stability.
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