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.

Science 26 September 2003:
Vol. 301. no. 5641, pp. 1921 - 1925
DOI: 10.1126/science.1088710
Prev | Table of Contents | Next

Reports

R2D2, a Bridge Between the Initiation and Effector Steps of the Drosophila RNAi Pathway

Qinghua Liu,1 Tim A. Rand,1 Savitha Kalidas,2 Fenghe Du,1 Hyun-Eui Kim,1 Dean P. Smith,2 Xiaodong Wang1*

The RNA interference (RNAi) pathway is initiated by processing long double-stranded RNA into small interfering RNA (siRNA). The siRNA-generating enzyme was purified from Drosophila S2cells and consists of two stoichiometric subunits: Dicer-2(DCR-2) and a previously unknown protein that we named R2D2. R2D2 is homologous to the Caenorhabditis elegans RNAi protein RDE-4. Association with R2D2 does not affect the enzymatic activity of DCR-2. Rather, the DCR-2/R2D2 complex, but not DCR-2 alone, binds to siRNA and enhances sequence-specific messenger RNA degradation mediated by the RNA-initiated silencing complex (RISC). These results indicate that R2D2 bridges the initiation and effector steps of the Drosophila RNAi pathway by facilitating siRNA passage from Dicer to RISC.

1 Howard Hughes Medical Institute and Department of Biochemistry, University of Texas (UT) Southwestern Medical Center at Dallas, Dallas, TX 75390, USA.
2 Department of Pharmacology and Center for Basic Neuroscience, University of Texas (UT) Southwestern Medical Center at Dallas, Dallas, TX 75390, USA.

* To whom correspondence should be addressed. E-mail: xwang{at}biochem.swmed.edu

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Sorting of Drosophila small silencing RNAs partitions microRNA* strands into the RNA interference pathway.
M. Ghildiyal, J. Xu, H. Seitz, Z. Weng, and P. D. Zamore (2010)
RNA 16, 43-56
   Abstract »    Full Text »    PDF »
The Arabidopsis thaliana double-stranded RNA binding protein DRB1 directs guide strand selection from microRNA duplexes.
A. L. Eamens, N. A. Smith, S. J. Curtin, M.-B. Wang, and P. M. Waterhouse (2009)
RNA 15, 2219-2235
   Abstract »    Full Text »    PDF »
RNAi in Budding Yeast.
I. A. Drinnenberg, D. E. Weinberg, K. T. Xie, J. P. Mower, K. H. Wolfe, G. R. Fink, and D. P. Bartel (2009)
Science 326, 544-550
   Abstract »    Full Text »    PDF »
Processing of Drosophila endo-siRNAs depends on a specific Loquacious isoform.
R. Zhou, B. Czech, J. Brennecke, R. Sachidanandam, J. A. Wohlschlegel, N. Perrimon, and G. J. Hannon (2009)
RNA 15, 1886-1895
   Abstract »    Full Text »    PDF »
C3PO, an Endoribonuclease That Promotes RNAi by Facilitating RISC Activation.
Y. Liu, X. Ye, F. Jiang, C. Liang, D. Chen, J. Peng, L. N. Kinch, N. V. Grishin, and Q. Liu (2009)
Science 325, 750-753
   Abstract »    Full Text »    PDF »
Characterization of the miRNA-RISC loading complex and miRNA-RISC formed in the Drosophila miRNA pathway.
K. Miyoshi, T. N. Okada, H. Siomi, and M. C. Siomi (2009)
RNA 15, 1282-1291
   Abstract »    Full Text »    PDF »
Genome-wide identification of targets of the drosha-pasha/DGCR8 complex.
S. Kadener, J. Rodriguez, K. C. Abruzzi, Y. L. Khodor, K. Sugino, M. T. Marr II, S. Nelson, and M. Rosbash (2009)
RNA 15, 537-545
   Abstract »    Full Text »    PDF »
Arabidopsis DRB4, AGO1, AGO7, and RDR6 participate in a DCL4-initiated antiviral RNA silencing pathway negatively regulated by DCL1.
F. Qu, X. Ye, and T. J. Morris (2008)
PNAS 105, 14732-14737
   Abstract »    Full Text »    PDF »
miRNAs: Effectors of Environmental Influences on Gene Expression and Disease.
A. Hudder and R. F. Novak (2008)
Toxicol. Sci. 103, 228-240
   Abstract »    Full Text »    PDF »
Functional dissection of siRNA sequence by systematic DNA substitution: modified siRNA with a DNA seed arm is a powerful tool for mammalian gene silencing with significantly reduced off-target effect.
K. Ui-Tei, Y. Naito, S. Zenno, K. Nishi, K. Yamato, F. Takahashi, A. Juni, and K. Saigo (2008)
Nucleic Acids Res. 36, 2136-2151
   Abstract »    Full Text »    PDF »
Identification of functional microRNAs released through asymmetrical processing of HIV-1 TAR element.
D. L. Ouellet, I. Plante, P. Landry, C. Barat, M.-E. Janelle, L. Flamand, M. J. Tremblay, and P. Provost (2008)
Nucleic Acids Res. 36, 2353-2365
   Abstract »    Full Text »    PDF »
aubergine Gene Overexpression in Somatic Tissues of auberginesting Mutants Interferes With the RNAi Pathway of a yellow Hairpin dsRNA in Drosophila melanogaster.
V. Specchia, C. Benna, G. M. Mazzotta, A. Piccin, M. A. Zordan, R. Costa, and M. P. Bozzetti (2008)
Genetics 178, 1271-1282
   Abstract »    Full Text »    PDF »
Structural and biochemical insights into the dicing mechanism of mouse Dicer: A conserved lysine is critical for dsRNA cleavage.
Z. Du, J. K. Lee, R. Tjhen, R. M. Stroud, and T. L. James (2008)
PNAS 105, 2391-2396
   Abstract »    Full Text »    PDF »
Strand-specific 5'-O-methylation of siRNA duplexes controls guide strand selection and targeting specificity.
P. Y. Chen, L. Weinmann, D. Gaidatzis, Y. Pei, M. Zavolan, T. Tuschl, and G. Meister (2008)
RNA 14, 263-274
   Abstract »    Full Text »    PDF »
Dicer-1, but not Loquacious, is critical for assembly of miRNA-induced silencing complexes.
X. Liu, J. K. Park, F. Jiang, Y. Liu, D. McKearin, and Q. Liu (2007)
RNA 13, 2324-2329
   Abstract »    Full Text »    PDF »
Evolutionary rate analyses of orthologs and paralogs from 12 Drosophila genomes.
A. Heger and C. P. Ponting (2007)
Genome Res. 17, 1837-1849
   Abstract »    Full Text »    PDF »
P68 RNA Helicase Unwinds the Human let-7 MicroRNA Precursor Duplex and Is Required for let-7-directed Silencing of Gene Expression.
D. W. Salzman, J. Shubert-Coleman, and H. Furneaux (2007)
J. Biol. Chem. 282, 32773-32779
   Abstract »    Full Text »    PDF »
Gene silencing mechanisms mediated by Aubergine piRNA complexes in Drosophila male gonad.
K. M. Nishida, K. Saito, T. Mori, Y. Kawamura, T. Nagami-Okada, S. Inagaki, H. Siomi, and M. C. Siomi (2007)
RNA 13, 1911-1922
   Abstract »    Full Text »    PDF »
Functional Anatomy of the Drosophila MicroRNA-generating Enzyme.
X. Ye, Z. Paroo, and Q. Liu (2007)
J. Biol. Chem. 282, 28373-28378
   Abstract »    Full Text »    PDF »
Depletion of newly synthesized Argonaute1 impairs the RNAi response in Trypanosoma brucei.
H. Shi, C. Tschudi, and E. Ullu (2007)
RNA 13, 1132-1139
   Abstract »    Full Text »    PDF »
Human TRBP and PACT Directly Interact with Each Other and Associate with Dicer to Facilitate the Production of Small Interfering RNA.
K. H. Kok, M.-H. J. Ng, Y.-P. Ching, and D.-Y. Jin (2007)
J. Biol. Chem. 282, 17649-17657
   Abstract »    Full Text »    PDF »
A Double-Stranded-RNA Response Program Important for RNA Interference Efficiency.
S. Choudhary, H.-C. Lee, M. Maiti, Q. He, P. Cheng, Q. Liu, and Y. Liu (2007)
Mol. Cell. Biol. 27, 3995-4005
   Abstract »    Full Text »    PDF »
QIP, a putative exonuclease, interacts with the Neurospora Argonaute protein and facilitates conversion of duplex siRNA into single strands.
M. Maiti, H.-C. Lee, and Y. Liu (2007)
Genes & Dev. 21, 590-600
   Abstract »    Full Text »    PDF »
A Novel Repeat-Associated Small Interfering RNA-Mediated Silencing Pathway Downregulates Complementary Sense gypsy Transcripts in Somatic Cells of the Drosophila Ovary.
A. Pelisson, E. Sarot, G. Payen-Groschene, and A. Bucheton (2007)
J. Virol. 81, 1951-1960
   Abstract »    Full Text »    PDF »
Conversion of pre-RISC to holo-RISC by Ago2 during assembly of RNAi complexes.
K. Kim, Y. S. Lee, and R. W. Carthew (2007)
RNA 13, 22-29
   Abstract »    Full Text »    PDF »
An unusual Dicer-like1 protein fuels the RNA interference pathway in Trypanosoma brucei.
H. Shi, C. Tschudi, and E. Ullu (2006)
RNA 12, 2063-2072
   Abstract »    Full Text »    PDF »
Molecular characterization of embryonic gonads by gene expression profiling in Drosophila melanogaster.
S. Shigenobu, Y. Kitadate, C. Noda, and S. Kobayashi (2006)
PNAS 103, 13728-13733
   Abstract »    Full Text »    PDF »
Characterization of DGCR8/Pasha, the essential cofactor for Drosha in primary miRNA processing.
K.-H. Yeom, Y. Lee, J. Han, M. R. Suh, and V. N. Kim (2006)
Nucleic Acids Res. 34, 4622-4629
   Abstract »    Full Text »    PDF »
Specific association of Piwi with rasiRNAs derived from retrotransposon and heterochromatic regions in the Drosophila genome.
K. Saito, K. M. Nishida, T. Mori, Y. Kawamura, K. Miyoshi, T. Nagami, H. Siomi, and M. C. Siomi (2006)
Genes & Dev. 20, 2214-2222
   Abstract »    Full Text »    PDF »
A genomewide screen for components of the RNAi pathway in Drosophila cultured cells.
S. Dorner, L. Lum, M. Kim, R. Paro, P. A. Beachy, and R. Green (2006)
PNAS 103, 11880-11885
   Abstract »    Full Text »    PDF »
Dicer-2 and R2D2 coordinately bind siRNA to promote assembly of the siRISC complexes.
X. Liu, F. Jiang, S. Kalidas, D. Smith, and Q. Liu (2006)
RNA 12, 1514-1520
   Abstract »    Full Text »    PDF »
A Distinct Small RNA Pathway Silences Selfish Genetic Elements in the Germline.
V. V. Vagin, A. Sigova, C. Li, H. Seitz, V. Gvozdev, and P. D. Zamore (2006)
Science 313, 320-324
   Abstract »    Full Text »    PDF »
Functional replacement of Trypanosoma brucei Argonaute by the human slicer Argonaute2.
H. Shi, C. Tschudi, and E. Ullu (2006)
RNA 12, 943-947
   Abstract »    Full Text »    PDF »
Double-stranded RNA Is Internalized by Scavenger Receptor-mediated Endocytosis in Drosophila S2 Cells.
J. Ulvila, M. Parikka, A. Kleino, R. Sormunen, R. A. Ezekowitz, C. Kocks, and M. Ramet (2006)
J. Biol. Chem. 281, 14370-14375
   Abstract »    Full Text »    PDF »
RDE-4 preferentially binds long dsRNA and its dimerization is necessary for cleavage of dsRNA to siRNA.
G. S. Parker, D. M. Eckert, and B. L. Bass (2006)
RNA 12, 807-818
   Abstract »    Full Text »    PDF »
RNA Interference Directs Innate Immunity Against Viruses in Adult Drosophila.
X.-H. Wang, R. Aliyari, W.-X. Li, H.-W. Li, K. Kim, R. Carthew, P. Atkinson, and S.-W. Ding (2006)
Science 312, 452-454
   Abstract »    Full Text »    PDF »
The interaction between DCL1 and HYL1 is important for efficient and precise processing of pri-miRNA in plant microRNA biogenesis.
Y. KURIHARA, Y. TAKASHI, and Y. WATANABE (2006)
RNA 12, 206-212
   Abstract »    Full Text »    PDF »
Structural Basis for Double-Stranded RNA Processing by Dicer.
I. J. MacRae, K. Zhou, F. Li, A. Repic, A. N. Brooks, W. Z. Cande, P. D. Adams, and J. A. Doudna (2006)
Science 311, 195-198
   Abstract »    Full Text »    PDF »
Profile of Xiaodong Wang.
N. Zagorski (2006)
PNAS 103, 7-9
   Full Text »    PDF »
The RNAi Pathway Initiated by Dicer-2 in Drosophila.
K. KIM, Y.S. LEE, D. HARRIS, K. NAKAHARA, and R.W. CARTHEW (2006)
Cold Spring Harb Symp Quant Biol 71, 39-44
   Abstract »    PDF »
Structure of Dicer and Mechanistic Implications for RNAi.
I.J. MacRAE, F. LI, K. ZHOU, W.Z. CANDE, and J.A. DOUDNA (2006)
Cold Spring Harb Symp Quant Biol 71, 73-80
   Abstract »    PDF »
Positive and Negative Modulation of Viral and Cellular mRNAs by Liver-specific MicroRNA miR-122.
C.L. JOPLING, K.L. NORMAN, and P. SARNOW (2006)
Cold Spring Harb Symp Quant Biol 71, 369-376
   Abstract »    PDF »
Small interfering RNAs containing full 2'-O-methylribonucleotide-modified sense strands display Argonaute2/eIF2C2-dependent activity.
B. A. KRAYNACK and B. F. BAKER (2006)
RNA 12, 163-176
   Abstract »    Full Text »    PDF »
Slicer function of Drosophila Argonautes and its involvement in RISC formation.
K. Miyoshi, H. Tsukumo, T. Nagami, H. Siomi, and M. C. Siomi (2005)
Genes & Dev. 19, 2837-2848
   Abstract »    Full Text »    PDF »
Molecular Requirements for RNA-induced Silencing Complex Assembly in the Drosophila RNA Interference Pathway.
J. W. Pham and E. J. Sontheimer (2005)
J. Biol. Chem. 280, 39278-39283
   Abstract »    Full Text »    PDF »
ATP modulates siRNA interactions with an endogenous human Dicer complex.
J. L. PELLINO, L. JASKIEWICZ, W. FILIPOWICZ, and E. J. SONTHEIMER (2005)
RNA 11, 1719-1724
   Abstract »    Full Text »    PDF »
A pathway for the biogenesis of trans-acting siRNAs in Arabidopsis.
M. Yoshikawa, A. Peragine, M. Y. Park, and R. S. Poethig (2005)
Genes & Dev. 19, 2164-2175
   Abstract »    Full Text »    PDF »
Characterization of Dicer-deficient murine embryonic stem cells.
E. P. Murchison, J. F. Partridge, O. H. Tam, S. Cheloufi, and G. J. Hannon (2005)
PNAS 102, 12135-12140
   Abstract »    Full Text »    PDF »
Functional polarity is introduced by Dicer processing of short substrate RNAs.
S. D. Rose, D.-H. Kim, M. Amarzguioui, J. D. Heidel, M. A. Collingwood, M. E. Davis, J. J. Rossi, and M. A. Behlke (2005)
Nucleic Acids Res. 33, 4140-4156
   Abstract »    Full Text »    PDF »
Dicer-1 and R3D1-L catalyze microRNA maturation in Drosophila.
F. Jiang, X. Ye, X. Liu, L. Fincher, D. McKearin, and Q. Liu (2005)
Genes & Dev. 19, 1674-1679
   Abstract »    Full Text »    PDF »
A potential link between transgene silencing and poly(A) tails.
M. C. SIOMI, H. TSUKUMO, A. ISHIZUKA, T. NAGAMI, and H. SIOMI (2005)
RNA 11, 1004-1011
   Abstract »    Full Text »    PDF »
A Virus-Encoded Inhibitor That Blocks RNA Interference in Mammalian Cells.
C. S. Sullivan and D. Ganem (2005)
J. Virol. 79, 7371-7379
   Abstract »    Full Text »    PDF »
In Vitro Synthesized Small Interfering RNAs Elicit RNA Interference in African Trypanosomes: AN IN VITRO AND IN VIVO ANALYSIS.
A. Best, L. Handoko, E. Schluter, and H. U. Goringer (2005)
J. Biol. Chem. 280, 20573-20579
   Abstract »    Full Text »    PDF »
MicroRNAs Regulate Brain Morphogenesis in Zebrafish.
A. J. Giraldez, R. M. Cinalli, M. E. Glasner, A. J. Enright, J. M. Thomson, S. Baskerville, S. M. Hammond, D. P. Bartel, and A. F. Schier (2005)
Science 308, 833-838
   Abstract »    Full Text »    PDF »
The contributions of dsRNA structure to Dicer specificity and efficiency.
A. VERMEULEN, L. BEHLEN, A. REYNOLDS, A. WOLFSON, W. S. MARSHALL, J. KARPILOW, and A. KHVOROVA (2005)
RNA 11, 674-682
   Abstract »    Full Text »    PDF »
An RNA polymerase II construct synthesizes short-hairpin RNA with a quantitative indicator and mediates highly efficient RNAi.
H. Zhou, X. G. Xia, and Z. Xu (2005)
Nucleic Acids Res. 33, e62
   Abstract »    Full Text »    PDF »
Perspective: machines for RNAi.
Y. Tomari and P. D. Zamore (2005)
Genes & Dev. 19, 517-529
   Abstract »    Full Text »    PDF »
Overexpression of Exportin 5 enhances RNA interference mediated by short hairpin RNAs and microRNAs.
R. YI, B. P. DOEHLE, Y. QIN, I. G. MACARA, and B. R. CULLEN (2005)
RNA 11, 220-226
   Abstract »    Full Text »    PDF »
RDE-2 interacts with MUT-7 to mediate RNA interference in Caenorhabditis elegans.
B. B. J. Tops, H. Tabara, T. Sijen, F. Simmer, C. C. Mello, R. H. A. Plasterk, and R. F. Ketting (2005)
Nucleic Acids Res. 33, 347-355
   Abstract »    Full Text »    PDF »
The Drosha-DGCR8 complex in primary microRNA processing.
J. Han, Y. Lee, K.-H. Yeom, Y.-K. Kim, H. Jin, and V. N. Kim (2004)
Genes & Dev. 18, 3016-3027
   Abstract »    Full Text »    PDF »
Selection and Characterization of RNA Interference-Deficient Trypanosomes Impaired in Target mRNA Degradation.
H. Shi, N. Chamond, C. Tschudi, and E. Ullu (2004)
Eukaryot. Cell 3, 1445-1453
   Abstract »    Full Text »    PDF »
A Protein Sensor for siRNA Asymmetry.
Y. Tomari, C. Matranga, B. Haley, N. Martinez, and P. D. Zamore (2004)
Science 306, 1377-1380
   Abstract »    Full Text »    PDF »
Biochemical identification of Argonaute 2 as the sole protein required for RNA-induced silencing complex activity.
T. A. Rand, K. Ginalski, N. V. Grishin, and X. Wang (2004)
PNAS 101, 14385-14389
   Abstract »    Full Text »    PDF »
RNA interference: from model organisms towards therapy for neural and neuromuscular disorders.
S. D. Buckingham, B. Esmaeili, M. Wood, and D. B. Sattelle (2004)
Hum. Mol. Genet. 13, R275-R288
   Abstract »    Full Text »    PDF »
Effects of Length and Location on the Cellular Response to Double-Stranded RNA.
Q. Wang and G. G. Carmichael (2004)
Microbiol. Mol. Biol. Rev. 68, 432-452
   Abstract »    Full Text »    PDF »
"siRNAs and miRNAs": A meeting report on RNA silencing.
Z. HE and E. J. SONTHEIMER (2004)
RNA 10, 1165-1173
   Full Text »    PDF »
Short Interfering RNA Accumulation Correlates with Host Recovery in DNA Virus-Infected Hosts, and Gene Silencing Targets Specific Viral Sequences.
P. Chellappan, R. Vanitharani, and C. M. Fauquet (2004)
J. Virol. 78, 7465-7477
   Abstract »    Full Text »    PDF »
Distinct roles for Argonaute proteins in small RNA-directed RNA cleavage pathways.
K. Okamura, A. Ishizuka, H. Siomi, and M. C. Siomi (2004)
Genes & Dev. 18, 1655-1666
   Abstract »    Full Text »    PDF »
Viral RNA silencing suppressors inhibit the microRNA pathway at an intermediate step.
E. J. Chapman, A. I. Prokhnevsky, K. Gopinath, V. V. Dolja, and J. C. Carrington (2004)
Genes & Dev. 18, 1179-1186
   Abstract »    Full Text »    PDF »
RISC is a 5' phosphomonoester-producing RNA endonuclease.
J. Martinez and T. Tuschl (2004)
Genes & Dev. 18, 975-980
   Abstract »    Full Text »    PDF »
The Arabidopsis double-stranded RNA-binding protein HYL1 plays a role in microRNA-mediated gene regulation.
M.-H. Han, S. Goud, L. Song, and N. Fedoroff (2004)
PNAS 101, 1093-1098
   Abstract »    Full Text »    PDF »
Plant RNA Interference in Vitro.
C. MATRANGA and P.D. ZAMORE (2004)
Cold Spring Harb Symp Quant Biol 69, 403-408
   Abstract »    PDF »
A Conserved microRNA Signal Specifies Leaf Polarity.
M.C.P. TIMMERMANS, M.T. JUAREZ, and T.L. PHELPS-DURR (2004)
Cold Spring Harb Symp Quant Biol 69, 409-418
   Abstract »    PDF »
The small RNA world.
E. J. Finnegan and M. A. Matzke (2003)
J. Cell Sci. 116, 4689-4693
   Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)