Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Subscribe

Logo for

Science 305 (5689): 1437-1441

Copyright © 2004 by the American Association for the Advancement of Science

Argonaute2 Is the Catalytic Engine of Mammalian RNAi

Jidong Liu,1* Michelle A. Carmell,1,2* Fabiola V. Rivas,1 Carolyn G. Marsden,1 J. Michael Thomson,3 Ji-Joon Song,1 Scott M. Hammond,3 Leemor Joshua-Tor,1 Gregory J. Hannon1{dagger}

Abstract: Gene silencing through RNA interference (RNAi) is carried out by RISC, the RNA-induced silencing complex. RISC contains two signature components, small interfering RNAs (siRNAs) and Argonaute family proteins. Here, we show that the multiple Argonaute proteins present in mammals are both biologically and biochemically distinct, with a single mammalian family member, Argonaute2, being responsible for messenger RNA cleavage activity. This protein is essential for mouse development, and cells lacking Argonaute2 are unable to mount an experimental response to siRNAs. Mutations within a cryptic ribonuclease H domain within Argonaute2, as identified by comparison with the structure of an archeal Argonaute protein, inactivate RISC. Thus, our evidence supports a model in which Argonaute contributes "Slicer" activity to RISC, providing the catalytic engine for RNAi.

1 Cold Spring Harbor Laboratory, Watson School of Biological Sciences, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.
2 Program in Genetics, Stony Brook University, Stony Brook, NY 11794, USA.
3 Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, NC 27599, USA.

Back to Top

* These authors contributed equally to this work.

{dagger} To whom correspondence should be addressed. E-mail: hannon{at}cshl.edu


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Elements and machinery of non-coding RNAs: toward their taxonomy.
T. Hirose, Y. Mishima, and Y. Tomari (2014)
EMBO Rep.
   Abstract »    Full Text »    PDF »
The RNase H-like superfamily: new members, comparative structural analysis and evolutionary classification.
K. A. Majorek, S. Dunin-Horkawicz, K. Steczkiewicz, A. Muszewska, M. Nowotny, K. Ginalski, and J. M. Bujnicki (2014)
Nucleic Acids Res. 42, 4160-4179
   Abstract »    Full Text »    PDF »
Traffic into silence: endomembranes and post-transcriptional RNA silencing.
Y. J. Kim, A. Maizel, and X. Chen (2014)
EMBO J.
   Abstract »    Full Text »    PDF »
Structure-based cleavage mechanism of Thermus thermophilus Argonaute DNA guide strand-mediated DNA target cleavage.
G. Sheng, H. Zhao, J. Wang, Y. Rao, W. Tian, D. C. Swarts, J. van der Oost, D. J. Patel, and Y. Wang (2014)
PNAS 111, 652-657
   Abstract »    Full Text »    PDF »
Computational prediction and experimental validation of microRNAs in the brown alga Ectocarpus siliculosus.
B. Billoud, Z. Nehr, A. Le Bail, and B. Charrier (2014)
Nucleic Acids Res. 42, 417-429
   Abstract »    Full Text »    PDF »
Enhancement of hepatitis C viral RNA abundance by precursor miR-122 molecules.
E. M. Cox, S. M. Sagan, S. A. W. Mortimer, J. A. Doudna, and P. Sarnow (2013)
RNA 19, 1825-1832
   Abstract »    Full Text »    PDF »
Self-duplexing CUG repeats selectively inhibit mutant huntingtin expression.
A. Fiszer, M. Olejniczak, P. Galka-Marciniak, A. Mykowska, and W. J. Krzyzosiak (2013)
Nucleic Acids Res. 41, 10426-10437
   Abstract »    Full Text »    PDF »
Promoter RNA links transcriptional regulation of inflammatory pathway genes.
M. Matsui, Y. Chu, H. Zhang, K. T. Gagnon, S. Shaikh, S. Kuchimanchi, M. Manoharan, D. R. Corey, and B. A. Janowski (2013)
Nucleic Acids Res. 41, 10086-10109
   Abstract »    Full Text »    PDF »
The co-chaperones Fkbp4/5 control Argonaute2 expression and facilitate RISC assembly.
N. J. Martinez, H.-M. Chang, J. d. R. Borrajo, and R. I. Gregory (2013)
RNA 19, 1583-1593
   Abstract »    Full Text »    PDF »
Robust RNAi enhancement via human Argonaute-2 overexpression from plasmids, viral vectors and cell lines.
K. Borner, D. Niopek, G. Cotugno, M. Kaldenbach, T. Pankert, J. Willemsen, X. Zhang, N. Schurmann, S. Mockenhaupt, A. Serva, et al. (2013)
Nucleic Acids Res. 41, e199
   Abstract »    Full Text »    PDF »
ss-siRNAs allele selectively inhibit ataxin-3 expression: multiple mechanisms for an alternative gene silencing strategy.
J. Liu, D. Yu, Y. Aiba, H. Pendergraff, E. E. Swayze, W. F. Lima, J. Hu, T. P. Prakash, and D. R. Corey (2013)
Nucleic Acids Res. 41, 9570-9583
   Abstract »    Full Text »    PDF »
Minimal mechanistic model of siRNA-dependent target RNA slicing by recombinant human Argonaute 2 protein.
A. Deerberg, S. Willkomm, and T. Restle (2013)
PNAS 110, 17850-17855
   Abstract »    Full Text »    PDF »
A component of the mir-17-92 polycistronic oncomir promotes oncogene-dependent apoptosis.
V. Olive, E. Sabio, M. J. Bennett, C. S. De Jong, A. Biton, J. C. McGann, S. K. Greaney, N. M. Sodir, A. Y. Zhou, A. Balakrishnan, et al. (2013)
eLife Sci 2, e00822
   Abstract »    Full Text »    PDF »
Core small nuclear ribonucleoprotein particle splicing factor SmD1 modulates RNA interference in Drosophila.
X.-P. Xiong, K. Kurthkoti, K.-Y. Chang, G. Lichinchi, N. De, A. Schneemann, I. J. MacRae, T. M. Rana, N. Perrimon, and R. Zhou (2013)
PNAS 110, 16520-16525
   Abstract »    Full Text »    PDF »
RNA duplexes with abasic substitutions are potent and allele-selective inhibitors of huntingtin and ataxin-3 expression.
J. Liu, H. Pendergraff, K. J. Narayanannair, J. G. Lackey, S. Kuchimanchi, K. G. Rajeev, M. Manoharan, J. Hu, and D. R. Corey (2013)
Nucleic Acids Res. 41, 8788-8801
   Abstract »    Full Text »    PDF »
Structural features of Argonaute-GW182 protein interactions.
J. Pfaff, J. Hennig, F. Herzog, R. Aebersold, M. Sattler, D. Niessing, and G. Meister (2013)
PNAS 110, E3770-E3779
   Abstract »    Full Text »    PDF »
VEGF controls lung Th2 inflammation via the miR-1-Mpl (myeloproliferative leukemia virus oncogene)-P-selectin axis.
S. Takyar, H. Vasavada, J.-g. Zhang, F. Ahangari, N. Niu, Q. Liu, C. G. Lee, L. Cohn, and J. A. Elias (2013)
J. Exp. Med. 210, 1993-2010
   Abstract »    Full Text »    PDF »
Hsp90 cochaperones p23 and FKBP4 physically interact with hAgo2 and activate RNA interference-mediated silencing in mammalian cells.
J. M. Pare, P. LaPointe, and T. C. Hobman (2013)
Mol. Biol. Cell 24, 2303-2310
   Abstract »    Full Text »    PDF »
Integrative Omics Analysis Reveals the Importance and Scope of Translational Repression in microRNA-mediated Regulation.
Q. Liu, P. J. Halvey, Y. Shyr, R. J. C. Slebos, D. C. Liebler, and B. Zhang (2013)
Mol. Cell. Proteomics 12, 1900-1911
   Abstract »    Full Text »    PDF »
Biogenesis, Turnover, and Mode of Action of Plant MicroRNAs.
K. Rogers and X. Chen (2013)
PLANT CELL 25, 2383-2399
   Abstract »    Full Text »    PDF »
Multiple sensors ensure guide strand selection in human RNAi pathways.
C. L. Noland and J. A. Doudna (2013)
RNA 19, 639-648
   Abstract »    Full Text »    PDF »
Argonaute2 expression is post-transcriptionally coupled to microRNA abundance.
N. J. Martinez and R. I. Gregory (2013)
RNA 19, 605-612
   Abstract »    Full Text »    PDF »
Loop-miRs: active microRNAs generated from single-stranded loop regions.
J. Winter, S. Link, D. Witzigmann, C. Hildenbrand, C. Previti, and S. Diederichs (2013)
Nucleic Acids Res. 41, 5503-5512
   Abstract »    Full Text »    PDF »
Argonaute2 Regulates the Pancreatic {beta}-Cell Secretome.
S. G. Tattikota, M. D. Sury, T. Rathjen, H.-H. Wessels, A. K. Pandey, X. You, C. Becker, W. Chen, M. Selbach, and M. N. Poy (2013)
Mol. Cell. Proteomics 12, 1214-1225
   Abstract »    Full Text »    PDF »
The rough endoplasmatic reticulum is a central nucleation site of siRNA-mediated RNA silencing.
L. Stalder, W. Heusermann, L. Sokol, D. Trojer, J. Wirz, J. Hean, A. Fritzsche, F. Aeschimann, V. Pfanzagl, P. Basselet, et al. (2013)
EMBO J. 32, 1115-1127
   Abstract »    Full Text »    PDF »
Context-specific microRNA function in developmental complexity.
A. P. Carroll, P. A. Tooney, and M. J. Cairns (2013)
J Mol Cell Biol 5, 73-84
   Abstract »    Full Text »    PDF »
Dicer-independent processing of short hairpin RNAs.
Y. P. Liu, N. C. T. Schopman, and B. Berkhout (2013)
Nucleic Acids Res. 41, 3723-3733
   Abstract »    Full Text »    PDF »
Functional small RNAs are generated from select miRNA hairpin loops in flies and mammals.
K. Okamura, E. Ladewig, L. Zhou, and E. C. Lai (2013)
Genes & Dev. 27, 778-792
   Abstract »    Full Text »    PDF »
Structural insights into RISC assembly facilitated by dsRNA-binding domains of human RNA helicase A (DHX9).
Q. Fu and Y. A. Yuan (2013)
Nucleic Acids Res. 41, 3457-3470
   Abstract »    Full Text »    PDF »
Mutations to the piRNA Pathway Component Aubergine Enhance Meiotic Drive of Segregation Distorter in Drosophila melanogaster.
S. L. Gell and R. A. Reenan (2013)
Genetics 193, 771-784
   Abstract »    Full Text »    PDF »
Circular RNAs are abundant, conserved, and associated with ALU repeats.
W. R. Jeck, J. A. Sorrentino, K. Wang, M. K. Slevin, C. E. Burd, J. Liu, W. F. Marzluff, and N. E. Sharpless (2013)
RNA 19, 141-157
   Abstract »    Full Text »    PDF »
Rapid and specific purification of Argonaute-small RNA complexes from crude cell lysates.
C. F. Flores-Jasso, W. E. Salomon, and P. D. Zamore (2013)
RNA 19, 271-279
   Abstract »    Full Text »    PDF »
Combinatorial mRNA binding by AUF1 and Argonaute 2 controls decay of selected target mRNAs.
X. Wu, S. Chesoni, G. Rondeau, C. Tempesta, R. Patel, S. Charles, N. Daginawala, B. E. Zucconi, A. Kishor, G. Xu, et al. (2013)
Nucleic Acids Res. 41, 2644-2658
   Abstract »    Full Text »    PDF »
tRNA-derived microRNA modulates proliferation and the DNA damage response and is down-regulated in B cell lymphoma.
R. L. Maute, C. Schneider, P. Sumazin, A. Holmes, A. Califano, K. Basso, and R. Dalla-Favera (2013)
PNAS 110, 1404-1409
   Abstract »    Full Text »    PDF »
Profiling of T helper cell-derived small RNAs reveals unique antisense transcripts and differential association of miRNAs with argonaute proteins 1 and 2.
S. Polikepahad and D. B. Corry (2013)
Nucleic Acids Res. 41, 1164-1177
   Abstract »    Full Text »    PDF »
Lessons on RNA Silencing Mechanisms in Plants from Eukaryotic Argonaute Structures.
C. Poulsen, H. Vaucheret, and P. Brodersen (2013)
PLANT CELL 25, 22-37
   Abstract »    Full Text »    PDF »
Mechanism of allele-selective inhibition of huntingtin expression by duplex RNAs that target CAG repeats: function through the RNAi pathway.
J. Hu, J. Liu, D. Yu, Y. Chu, and D. R. Corey (2012)
Nucleic Acids Res. 40, 11270-11280
   Abstract »    Full Text »    PDF »
Alternative RISC assembly: Binding and repression of microRNA-mRNA duplexes by human Ago proteins.
M. M. Janas, B. Wang, A. S. Harris, M. Aguiar, J. M. Shaffer, Y. V. B. K. Subrahmanyam, M. A. Behlke, K. W. Wucherpfennig, S. P. Gygi, E. Gagnon, et al. (2012)
RNA 18, 2041-2055
   Abstract »    Full Text »    PDF »
The slicing activity of miRNA-specific Argonautes is essential for the miRNA pathway in C. elegans.
S. Bouasker and M. J. Simard (2012)
Nucleic Acids Res. 40, 10452-10462
   Abstract »    Full Text »    PDF »
HIV-1 Replication and APOBEC3 Antiviral Activity Are Not Regulated by P Bodies.
P. K. Phalora, N. M. Sherer, S. M. Wolinsky, C. M. Swanson, and M. H. Malim (2012)
J. Virol. 86, 11712-11724
   Abstract »    Full Text »    PDF »
Dicer-dependent and -independent Argonaute2 Protein Interaction Networks in Mammalian Cells.
A. Frohn, H. C. Eberl, J. Stohr, E. Glasmacher, S. Rudel, V. Heissmeyer, M. Mann, and G. Meister (2012)
Mol. Cell. Proteomics 11, 1442-1456
   Abstract »    Full Text »    PDF »
Right- and left-loop short shRNAs have distinct and unusual mechanisms of gene silencing.
A. Dallas, H. Ilves, Q. Ge, P. Kumar, J. Shorenstein, S. A. Kazakov, T. L. Cuellar, M. T. McManus, M. A. Behlke, and B. H. Johnston (2012)
Nucleic Acids Res. 40, 9255-9271
   Abstract »    Full Text »    PDF »
microRNAs associated with the different human Argonaute proteins.
A. Dueck, C. Ziegler, A. Eichner, E. Berezikov, and G. Meister (2012)
Nucleic Acids Res. 40, 9850-9862
   Abstract »    Full Text »    PDF »
N-myc and Noncoding RNAs in Neuroblastoma.
J. Buechner and C. Einvik (2012)
Mol. Cancer Res. 10, 1243-1253
   Abstract »    Full Text »    PDF »
APOBEC3G Inhibits MicroRNA-mediated Repression of Translation by Interfering with the Interaction between Argonaute-2 and MOV10.
C. Liu, X. Zhang, F. Huang, B. Yang, J. Li, B. Liu, H. Luo, P. Zhang, and H. Zhang (2012)
J. Biol. Chem. 287, 29373-29383
   Abstract »    Full Text »    PDF »
Mammalian GW220/TNGW1 is essential for the formation of GW/P bodies containing miRISC.
V. Castilla-Llorente, L. Spraggon, M. Okamura, S. Naseeruddin, M. Adamow, S. Qamar, and J. Liu (2012)
J. Cell Biol. 198, 529-544
   Abstract »    Full Text »    PDF »
Extensive terminal and asymmetric processing of small RNAs from rRNAs, snoRNAs, snRNAs, and tRNAs.
Z. Li, C. Ender, G. Meister, P. S. Moore, Y. Chang, and B. John (2012)
Nucleic Acids Res. 40, 6787-6799
   Abstract »    Full Text »    PDF »
Argonaute identity defines the length of mature mammalian microRNAs.
P. K. Juvvuna, P. Khandelia, L. M. Lee, and E. V. Makeyev (2012)
Nucleic Acids Res. 40, 6808-6820
   Abstract »    Full Text »    PDF »
Murine cytomegalovirus infection of cultured mouse cells induces expression of miR-7a.
A. Dittmer and K. Forstemann (2012)
J. Gen. Virol. 93, 1537-1547
   Abstract »    Full Text »    PDF »
HuR protein attenuates miRNA-mediated repression by promoting miRISC dissociation from the target RNA.
P. Kundu, M. R. Fabian, N. Sonenberg, S. N. Bhattacharyya, and W. Filipowicz (2012)
Nucleic Acids Res. 40, 5088-5100
   Abstract »    Full Text »    PDF »
The Crystal Structure of Human Argonaute2.
N. T. Schirle and I. J. MacRae (2012)
Science 336, 1037-1040
   Abstract »    Full Text »    PDF »
MicroRNA Targeting of Neurotropic Flavivirus: Effective Control of Virus Escape and Reversion to Neurovirulent Phenotype.
B. L. Heiss, O. A. Maximova, D. C. Thach, J. M. Speicher, and A. G. Pletnev (2012)
J. Virol. 86, 5647-5659
   Abstract »    Full Text »    PDF »
Functional parameters of Dicer-independent microRNA biogenesis.
J.-S. Yang, T. Maurin, and E. C. Lai (2012)
RNA 18, 945-957
   Abstract »    Full Text »    PDF »
Biogenesis of mammalian microRNAs by a non-canonical processing pathway.
M. A. Havens, A. A. Reich, D. M. Duelli, and M. L. Hastings (2012)
Nucleic Acids Res. 40, 4626-4640
   Abstract »    Full Text »    PDF »
Enhanced Susceptibility of Ago1/3 Double-Null Mice to Influenza A Virus Infection.
M. Van Stry, T. H. Oguin III, S. Cheloufi, P. Vogel, M. Watanabe, M. R. Pillai, P. Dash, P. G. Thomas, G. J. Hannon, and M. Bix (2012)
J. Virol. 86, 4151-4157
   Abstract »    Full Text »    PDF »
The RDE-10/RDE-11 complex triggers RNAi-induced mRNA degradation by association with target mRNA in C. elegans.
H. Yang, Y. Zhang, J. Vallandingham, H. Li, L. Florens, and H. Y. Mak (2012)
Genes & Dev. 26, 846-856
   Abstract »    Full Text »    PDF »
Quantitative functions of Argonaute proteins in mammalian development.
D. Wang, Z. Zhang, E. O'Loughlin, T. Lee, S. Houel, D. O'Carroll, A. Tarakhovsky, N. G. Ahn, and R. Yi (2012)
Genes & Dev. 26, 693-704
   Abstract »    Full Text »    PDF »
Expression determinants of mammalian argonaute proteins in mediating gene silencing.
P. N. Valdmanis, S. Gu, N. Schuermann, P. Sethupathy, D. Grimm, and M. A. Kay (2012)
Nucleic Acids Res. 40, 3704-3713
   Abstract »    Full Text »    PDF »
Widespread RNA 3'-end oligouridylation in mammals.
Y. S. Choi, W. Patena, A. D. Leavitt, and M. T. McManus (2012)
RNA 18, 394-401
   Abstract »    Full Text »    PDF »
MECHANISMS IN ENDOCRINOLOGY: Micro-RNAs: targets for enhancing osteoblast differentiation and bone formation.
H. Taipaleenmaki, L. Bjerre Hokland, L. Chen, S. Kauppinen, and M. Kassem (2012)
Eur. J. Endocrinol. 166, 359-371
   Abstract »    Full Text »    PDF »
Trinucleotide Repeat Containing 6a (Tnrc6a)-mediated MicroRNA Function Is Required for Development of Yolk Sac Endoderm.
Z. Jiang, N. Yu, P. Kuang, M. Chen, F. Shao, G. Martin, D. H. K. Chui, W. V. Cardoso, X. Ai, and J. Lu (2012)
J. Biol. Chem. 287, 5979-5987
   Abstract »    Full Text »    PDF »
Strand antagonism in RNAi: an explanation of differences in potency between intracellularly expressed siRNA and shRNA.
X. Jin, T. Sun, C. Zhao, Y. Zheng, Y. Zhang, W. Cai, Q. He, K. Taira, L. Zhang, and D. Zhou (2012)
Nucleic Acids Res. 40, 1797-1806
   Abstract »    Full Text »    PDF »
Expanding the action of duplex RNAs into the nucleus: redirecting alternative splicing.
J. Liu, J. Hu, and D. R. Corey (2012)
Nucleic Acids Res. 40, 1240-1250
   Abstract »    Full Text »    PDF »
Direct evidence of nuclear Argonaute distribution during transcriptional silencing links the actin cytoskeleton to nuclear RNAi machinery in human cells.
C. L. Ahlenstiel, H. G. W. Lim, D. A. Cooper, T. Ishida, A. D. Kelleher, and K. Suzuki (2012)
Nucleic Acids Res. 40, 1579-1595
   Abstract »    Full Text »    PDF »
Dicer is dispensable for asymmetric RISC loading in mammals.
J. G. Betancur and Y. Tomari (2012)
RNA 18, 24-30
   Abstract »    Full Text »    PDF »
microRNA Biogenesis and Turnover in Plants.
K. Rogers and X. Chen (2012)
Cold Spring Harb Symp Quant Biol 77, 183-194
   Abstract »    Full Text »    PDF »
Different Mutations in Three Prime Repair Exonuclease 1 and Ribonuclease H2 Genes Affect Clinical Features in Aicardi-Goutieres Syndrome.
A. Izzotti, M. Longobardi, C. Cartiglia, F. Anzuini, P. Arrigo, E. Fazzi, S. Orcesi, R. L. Piana, and A. Pulliero (2012)
J Child Neurol 27, 51-60
   Abstract »    Full Text »    PDF »
miRNA and mammalian male germ cells.
S. C. McIver, S. D. Roman, B. Nixon, and E. A. McLaughlin (2012)
Hum. Reprod. Update 18, 44-59
   Abstract »    Full Text »    PDF »
A microRNA derived from an apparent canonical biogenesis pathway regulates variant surface protein gene expression in Giardia lamblia.
A. A. Saraiya, W. Li, and C. C. Wang (2011)
RNA 17, 2152-2164
   Abstract »    Full Text »    PDF »
Green tea polyphenol EGCG suppresses lung cancer cell growth through upregulating miR-210 expression caused by stabilizing HIF-1{alpha}.
H. Wang, S. Bian, and C. S. Yang (2011)
Carcinogenesis 32, 1881-1889
   Abstract »    Full Text »    PDF »
Binding of the Cyclophilin 40 Ortholog SQUINT to Hsp90 Protein Is Required for SQUINT Function in Arabidopsis.
K. W. Earley and R. S. Poethig (2011)
J. Biol. Chem. 286, 38184-38189
   Abstract »    Full Text »    PDF »
miRNA-dependent gene silencing involving Ago2-mediated cleavage of a circular antisense RNA.
T. B. Hansen, E. D. Wiklund, J. B. Bramsen, S. B. Villadsen, A. L. Statham, S. J. Clark, and J. Kjems (2011)
EMBO J. 30, 4414-4422
   Abstract »    Full Text »    PDF »

To Advertise     Find Products


Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882