Prolyl 4-hydroxylation regulates Argonaute 2 stability.
Prolyl 4-hydroxylation regulates Argonaute 2 stability.
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DOI：
10.1038/nature07186
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发表时间：
2008-09-18
期刊：
影响因子：
64.8
通讯作者：
Shi, Yang
中科院分区：
文献类型：
--
作者：
Qi, Hank H.;Ongusaha, Pat P.;Myllyharju, Johanna;Cheng, Dongmei;Pakkanen, Outi;Shi, Yujiang;Lee, Sam W.;Peng, Junmin;Shi, Yang
人Argonaute（Ago）蛋白是RNA诱导沉默复合物（RISC）的重要组成部分。Argonaute 2（Ago 2）具有P元件诱导的懦弱睾丸（PIWI）结构域，其像RNA酶H一样折叠，并负责RNA干扰中的靶RNA切割。蛋白质如Dicer、TRBP、M0 V10、RHA、RCK/p54和KIAA 1093与Ago蛋白质缔合并参与小RNA加工、RISC装载和Ago蛋白质在细胞质信使RNA加工体中的定位。然而，调节RNA干扰的机制仍然不清楚。本文报道了Ago 2与I型胶原脯氨酰-4-羟化酶（C-P4 H（I））α-（P4 H-α（I））和β-（P4 H-β）亚基之间的物理相互作用。质谱分析鉴定了内源性Ago 2在脯氨酸700处的羟基化。在体外，Ago 2和Ago 4似乎比Ago 1和Ago 3更有效地被重组人C-P4 H（I）羟基化。重要的是，通过短发夹RNA去除P4 H-α（I）或P4 H-β的人细胞和P4 H-α（I）缺失的小鼠胚胎成纤维细胞显示Ago 2稳定性降低和短干扰RNA程序化RISC活性受损。此外，脯氨酸700突变为丙氨酸也导致Ago 2的不稳定，从而将Ago 2 P700和该残基处的羟基化与其稳定性调节联系起来。这些发现将羟基化鉴定为对Ago 2稳定性和有效RNA干扰重要的翻译后修饰。
Human Argonaute (Ago) proteins are essential components of the RNA-induced silencing complexes (RISCs). Argonaute 2 (Ago2) has a P-element-induced wimpy testis (PIWI) domain, which folds like RNase H and is responsible for target RNA cleavage in RNA interference. Proteins such as Dicer, TRBP, MOV10, RHA, RCK/p54 and KIAA1093 associate with Ago proteins and participate in small RNA processing, RISC loading and localization of Ago proteins in the cytoplasmic messenger RNA processing bodies. However, mechanisms that regulate RNA interference remain obscure. Here we report physical interactions between Ago2 and the α-(P4H-α(I)) and β-(P4H-β) subunits of the type I collagen prolyl-4-hydroxylase (C-P4H(I)). Mass spectrometric analysis identified hydroxylation of the endogenous Ago2 at proline 700. In vitro, both Ago2 and Ago4 seem to be more efficiently hydroxylated than Ago1 and Ago3 by recombinant human C-P4H(I). Importantly, human cells depleted of P4H-α(I) or P4H-β by short hairpin RNA and P4H-α(I) null mouse embryonic fibroblast cells showed reduced stability of Ago2 and impaired short interfering RNA programmed RISC activity. Furthermore, mutation of proline 700 to alanine also resulted in destabilization of Ago2, thus linking Ago2 P700 and hydroxylation at this residue to its stability regulation. These findings identify hydroxylation as a post-translational modification important for Ago2 stability and effective RNA interference.
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DOI：
10.1046/j.1432-1033.2003.03846.x
发表时间：
2003-11-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Hofbauer, KH;Gess, B;Kurtz, A
通讯作者：
Kurtz, A
影响因子：
4.8
作者：
Holster, Tiina;Pakkanen, Outi;Myllyharju, Johanna
通讯作者：
Myllyharju, Johanna
DOI：
10.1073/pnas.89.16.7467
发表时间：
1992-08-15
影响因子：
11.1
作者：
VUORI, K;PIHLAJANIEMI, T;KIVIRIKKO, KI
通讯作者：
KIVIRIKKO, KI
影响因子：
21.3
作者：
Sen, GL;Blau, HM
通讯作者：
Blau, HM
影响因子：
64.5
作者：
Grishok, A;Pasquinelli, AE;Mello, CC
通讯作者：
Mello, CC