5'-mRNA-帽与人类 AGO2 MID 结构域相互作用的结构分析。
Structural analysis of 5'-mRNA-cap interactions with the human AGO2 MID domain.
机构信息
Department of Biochemistry, McGill University, 1160 Pine Avenue West, Montreal, Quebec H3A 1A3, Canada.
出版信息
EMBO Rep. 2011 May;12(5):415-20. doi: 10.1038/embor.2011.48. Epub 2011 Apr 8.
Abstract
In RNA silencing, microRNA (miRNA)-mediated translational repression occurs through mechanisms that do not invoke messenger-RNA (mRNA) target cleavage by Argonaute proteins. The nature of these mechanisms is unclear, but several recent studies have proposed that a direct interaction between the mRNA-cap and the middle (MID) domain of Argonautes is involved. Here, we present crystallographic and NMR data demonstrating that cap analogues do not bind significantly to the isolated MID domain of human Argonaute 2 (hAGO2) and are found in the miRNA 5'-nucleotide binding site in an implausible binding mode. Additionally, in vitro pull-down experiments with full-length hAGO2 indicate that the interaction with cap analogues is nonspecific.
摘要
在 RNA 沉默中，miRNA（微 RNA）介导的翻译抑制是通过不依赖 Argonaute 蛋白切割信使 RNA（mRNA）靶标的机制发生的。这些机制的性质尚不清楚，但最近的几项研究提出，mRNA 帽和 Argonautes 的中间（MID）结构域之间的直接相互作用涉及其中。在这里，我们展示了晶体学和 NMR 数据，证明帽类似物与分离的人 Argonaute 2（hAGO2）的中间结构域没有显著结合，并且以不合理的结合模式存在于 miRNA 5'-核苷酸结合位点中。此外，全长 hAGO2 的体外下拉实验表明，与帽类似物的相互作用是非特异性的。
相似文献
1
Structural analysis of 5'-mRNA-cap interactions with the human AGO2 MID domain.
EMBO Rep. 2011 May;12(5):415-20. doi: 10.1038/embor.2011.48. Epub 2011 Apr 8.
2
The human Ago2 MC region does not contain an eIF4E-like mRNA cap binding motif.
Biol Direct. 2009 Jan 21;4:2. doi: 10.1186/1745-6150-4-2.
3
An mRNA m7G cap binding-like motif within human Ago2 represses translation.
Cell. 2007 Jun 15;129(6):1141-51. doi: 10.1016/j.cell.2007.05.016. Epub 2007 May 24.
4
eIF4GI facilitates the MicroRNA-mediated gene silencing.
PLoS One. 2013;8(2):e55725. doi: 10.1371/journal.pone.0055725. Epub 2013 Feb 7.
5
Allosteric regulation of Argonaute proteins by miRNAs.
Nat Struct Mol Biol. 2010 Feb;17(2):144-50. doi: 10.1038/nsmb.1736. Epub 2010 Jan 10.
6
Let-7 microRNA-mediated mRNA deadenylation and translational repression in a mammalian cell-free system.
Genes Dev. 2007 Aug 1;21(15):1857-62. doi: 10.1101/gad.1566707.
7
Structural basis for 5'-nucleotide base-specific recognition of guide RNA by human AGO2.
Nature. 2010 Jun 10;465(7299):818-22. doi: 10.1038/nature09039. Epub 2010 May 26.
8
Mammalian GW182 contains multiple Argonaute-binding sites and functions in microRNA-mediated translational repression.
RNA. 2009 Jun;15(6):1078-89. doi: 10.1261/rna.1363109. Epub 2009 Apr 27.
9
The structure of human argonaute-2 in complex with miR-20a.
Cell. 2012 Jul 6;150(1):100-10. doi: 10.1016/j.cell.2012.05.017. Epub 2012 Jun 7.
10
Ago2/miRISC-mediated inhibition of CBP80/20-dependent translation and thereby abrogation of nonsense-mediated mRNA decay require the cap-associating activity of Ago2.
FEBS Lett. 2011 Sep 2;585(17):2682-7. doi: 10.1016/j.febslet.2011.07.047. Epub 2011 Aug 10.
引用本文的文献
1
Investigation of Strand-Selective Interaction of SNA-Modified siRNA with AGO2-MID.
Int J Mol Sci. 2020 Jul 23;21(15):5218. doi: 10.3390/ijms21155218.
2
Mechanistic Insights into MicroRNA-Mediated Gene Silencing.
Cold Spring Harb Perspect Biol. 2019 Mar 1;11(3):a032771. doi: 10.1101/cshperspect.a032771.
3
Differences in silencing of mismatched targets by sliced versus diced siRNAs.
Nucleic Acids Res. 2018 Jul 27;46(13):6806-6822. doi: 10.1093/nar/gky287.
4
A Simple and Cost-Effective Approach for In Vitro Production of Sliced siRNAs as Potent Triggers for RNAi.
Mol Ther Nucleic Acids. 2017 Sep 15;8:345-355. doi: 10.1016/j.omtn.2017.07.008. Epub 2017 Jul 13.
5
MicroRNAs recruit eIF4E2 to repress translation of target mRNAs.
Protein Cell. 2017 Oct;8(10):750-761. doi: 10.1007/s13238-017-0444-0. Epub 2017 Jul 28.
6
siRNA carrying an (E)-vinylphosphonate moiety at the 5΄ end of the guide strand augments gene silencing by enhanced binding to human Argonaute-2.
Nucleic Acids Res. 2017 Apr 7;45(6):3528-3536. doi: 10.1093/nar/gkw1171.
7
MicroRNA-mediated target mRNA cleavage and 3'-uridylation in human cells.
Sci Rep. 2016 Jul 21;6:30242. doi: 10.1038/srep30242.
8
Guidelines for the optimal design of miRNA-based shRNAs.
Methods. 2016 Jul 1;103:157-66. doi: 10.1016/j.ymeth.2016.04.003. Epub 2016 Apr 12.
9
miRISC recruits decapping factors to miRNA targets to enhance their degradation.
Nucleic Acids Res. 2013 Oct;41(18):8692-705. doi: 10.1093/nar/gkt619. Epub 2013 Jul 17.
10
Molecular dissection of human Argonaute proteins by DNA shuffling.
Nat Struct Mol Biol. 2013 Jul;20(7):818-26. doi: 10.1038/nsmb.2607. Epub 2013 Jun 9.
本文引用的文献
1
Processing of X-ray diffraction data collected in oscillation mode.
Methods Enzymol. 1997;276:307-26. doi: 10.1016/S0076-6879(97)76066-X.
2
Mammalian microRNAs predominantly act to decrease target mRNA levels.
Nature. 2010 Aug 12;466(7308):835-40. doi: 10.1038/nature09267.
3
Crystal structure and ligand binding of the MID domain of a eukaryotic Argonaute protein.
EMBO Rep. 2010 Jul;11(7):522-7. doi: 10.1038/embor.2010.81. Epub 2010 Jun 11.
4
Regulation of mRNA translation and stability by microRNAs.
Annu Rev Biochem. 2010;79:351-79. doi: 10.1146/annurev-biochem-060308-103103.
5
Structural basis for 5'-nucleotide base-specific recognition of guide RNA by human AGO2.
Nature. 2010 Jun 10;465(7299):818-22. doi: 10.1038/nature09039. Epub 2010 May 26.
6
PHENIX: a comprehensive Python-based system for macromolecular structure solution.
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21. doi: 10.1107/S0907444909052925. Epub 2010 Jan 22.
7
Allosteric regulation of Argonaute proteins by miRNAs.
Nat Struct Mol Biol. 2010 Feb;17(2):144-50. doi: 10.1038/nsmb.1736. Epub 2010 Jan 10.
8
Mechanisms of miRNA-mediated post-transcriptional regulation in animal cells.
Curr Opin Cell Biol. 2009 Jun;21(3):452-60. doi: 10.1016/j.ceb.2009.04.009. Epub 2009 May 18.
9
Drosophila argonaute1 and argonaute2 employ distinct mechanisms for translational repression.
Mol Cell. 2009 Apr 10;34(1):58-67. doi: 10.1016/j.molcel.2009.02.010. Epub 2009 Mar 5.
10
MicroRNAs: target recognition and regulatory functions.
Cell. 2009 Jan 23;136(2):215-33. doi: 10.1016/j.cell.2009.01.002.
Zotero 插件