Nuclear RNA Exosome at 3.1 Å Reveals Substrate Specificities, RNA Paths, and Allosteric Inhibition of Rrp44/Dis3.
Nuclear RNA Exosome at 3.1 Å Reveals Substrate Specificities, RNA Paths, and Allosteric Inhibition of Rrp44/Dis3.
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DOI：
10.1016/j.molcel.2016.09.038
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发表时间：
2016-11-17
期刊：
影响因子：
16
通讯作者：
Lima, Christopher D.
中科院分区：
文献类型：
--
作者：
Zinder, John C.;Wasmuth, Elizabeth V.;Lima, Christopher D.
真核生物RNA外泌体是一种必需的、保守的3′至5′外核糖核酸酶复合物，它能降解或处理几乎所有类型的细胞RNA。核RNA外泌体包括一个9个亚基的非催化核心，它结合Rrp44 （Dis3）和Rrp6亚基，分别调节它们的过程性和分布性3 ‘至5 ’外核糖核酸酶活性。在这里，我们利用具有两个3 '端的工程RNA获得了11亚基核外显体在3.1 Å与RNA结合的晶体结构。该结构揭示了延伸到Rrp6的RNA路径，该路径可穿透非催化核心，非催化核心与Rrp44之间的接触可抑制外核糖核酸酶活性，以及Rrp44外核糖核酸酶位点支持其降解3 '磷酸核糖核酸底物的能力的特征。利用重组的外泌体复合物，我们发现3 '磷酸核糖核酸不是Rrp6的底物，但在核外泌体中很容易被Rrp44降解。Zinder等人使用工程化的3 ' -3 ' RNA获得酵母11亚基核RNA外显体的高分辨率结构，RNA与其外核糖核酸酶Rrp6和Rrp44结合。该结构揭示了Rrp44和Rrp6的底物特异性，以及调节其活性的9个亚基非催化核心的特征。
The eukaryotic RNA exosome is an essential and conserved 3’ to 5’ exoribonuclease complex that degrades or processes nearly every class of cellular RNA. The nuclear RNA exosome includes a nine-subunit non-catalytic core that binds Rrp44 (Dis3) and Rrp6 subunits to modulate their processive and distributive 3’ to 5’ exoribonuclease activities, respectively. Here, we utilize an engineered RNA with two 3’ ends to obtain a crystal structure of an eleven-subunit nuclear exosome bound to RNA at 3.1 Å. The structure reveals an extended RNA path to Rrp6 that penetrates into the non-catalytic core, contacts between the non-catalytic core and Rrp44 that inhibit exoribonuclease activity, and features of the Rrp44 exoribonuclease site that support its ability to degrade 3’ phosphate RNA substrates. Using reconstituted exosome complexes, we show that 3’ phosphate RNA is not a substrate for Rrp6, but is readily degraded by Rrp44 in the nuclear exosome. Zinder et al. use an engineered 3’-3’ RNA to obtain a high-resolution structure of the yeast eleven-subunit nuclear RNA exosome with RNA bound to its exoribonucleases, Rrp6 and Rrp44. The structure reveals substrate specificities for Rrp44 and Rrp6 and features of the nine-subunit non-catalytic core that modulate their activities.
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影响因子：
14.9
作者：
Drazkowska K;Tomecki R;Stodus K;Kowalska K;Czarnocki-Cieciura M;Dziembowski A
通讯作者：
Dziembowski A
影响因子：
16
作者：
Kowalinski E;Kögel A;Ebert J;Reichelt P;Stegmann E;Habermann B;Conti E
通讯作者：
Conti E
DOI：
10.1261/rna.057760.116
发表时间：
2016-09
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Domanski M;Upla P;Rice WJ;Molloy KR;Ketaren NE;Stokes DL;Jensen TH;Rout MP;LaCava J
通讯作者：
LaCava J
影响因子：
10.5
作者：
Allmang, C;Petfalski, E;Mitchell, P
通讯作者：
Mitchell, P
影响因子：
64.5
作者：
Liu, Quansheng;Greimann, Jaclyn C.;Lima, Christopher D.
通讯作者：
Lima, Christopher D.