Argonaute2 Is the Catalytic Engine of Mammalian RNAi
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.
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References and Notes
1
G. J. Hannon, Nature418, 244 (2002).
2
A. Fireet al., Nature391, 806 (1998).
3
G. Hutvagner, P. D. Zamore, Curr. Opin. Genet. Dev.12, 225 (2002).
4
A. Hamilton, O. Voinnet, L. Chappell, D. Baulcombe, EMBO J.21, 4671 (2002).
5
E. Bernstein, A. A. Caudy, S. M. Hammond, G. J. Hannon, Nature409, 363 (2001).
6
Y. Leeet al., Nature425, 415 (2003).
7
S. M. Hammond, E. Bernstein, D. Beach, G. J. Hannon, Nature404, 293 (2000).
8
M. F. Mette, W. Aufsatz, J. van der Winden, M. A. Matzke, A. J. Matzke, EMBO J.19, 5194 (2000).
9
I. M. Hallet al., Science297, 2232 (2002).
10
T. Volpeet al., Science297, 1833 (2002).
11
M. Pal-Bhadra, U. Bhadra, J. A. Birchler, Mol. Cell9, 315 (2002).
12
P. H. Olsen, V. Ambros, Dev. Biol.216, 671 (1999).
13
D. P. Bartel, Cell116, 281 (2004).
14
T. Tuschl, P. D. Zamore, R. Lehmann, D. P. Bartel, P. A. Sharp, Genes Dev.13, 3191 (1999).
15
P. D. Zamore, T. Tuschl, P. A. Sharp, D. P. Bartel, Cell101, 25 (2000).
16
S. M. Elbashir, J. Martinez, A. Patkaniowska, W. Lendeckel, T. Tuschl, EMBO J.20, 6877 (2001).
17
J. Martinez, A. Patkaniowska, H. Urlaub, R. Luhrmann, T. Tuschl, Cell110, 563 (2002).
18
G. Hutvagner, P. D. Zamore, Science297, 2056 (2002).
19
S. M. Hammond, S. Boettcher, A. A. Caudy, R. Kobayashi, G. J. Hannon, Science293, 1146 (2001).
20
M. A. Carmell, G. J. Hannon, Nature Struct. Mol. Biol.11, 214 (2004).
21
L. Cerutti, N. Mian, A. Bateman, Trends Biochem. Sci.25, 481 (2000).
22
H. Tabaraet al., Cell99, 123 (1999).
23
M. A. Carmell, Z. Xuan, M. Q. Zhang, G. J. Hannon, Genes Dev.16, 2733 (2002).
24
A. A. Caudy, M. Myers, G. J. Hannon, S. M. Hammond, Genes Dev.16, 2491 (2002).
25
K. Okamura, A. Ishizuka, H. Siomi, M. C. Siomi, Genes Dev.18, 1655 (2004).
26
B. Zheng, A. A. Mills, A. Bradley, Nucleic Acids Res.27, 2354 (1999).
27
S. J. Conway, A. Kruzynska-Frejtag, P. L. Kneer, M. Machnicki, S. V. Koushik, Genesis35, 1 (2003).
28
W. Deng, H. Lin, Dev. Cell2, 819 (2002).
29
S. Kuramochi-Miyagawaet al., Development131, 839 (2004).
30
T. Sasaki, A. Shiohama, S. Minoshima, N. Shimizu, Genomics82, 323 (2003).
31
S. Yekta, I. H. Shih, D. P. Bartel, Science304, 594 (2004).
32
J. G. Doench, C. P. Petersen, P. A. Sharp, Genes Dev.17, 438 (2003).
33
M. Kiriakidouet al., Genes Dev.18, 1165 (2004).
34
A. Nykanen, B. Haley, P. D. Zamore, Cell107, 309 (2001).
35
J. W. Pham, J. L. Pellino, Y. S. Lee, R. W. Carthew, E. J. Sontheimer, Cell117, 83 (2004).
36
Y. Tomariet al., Cell116, 831 (2004).
37
J.-J. Song et al., Science305, 1434 (2004). Published online 29 July 2004; 10.1126/science.1102514.
38
B. R. Chapadoset al., J. Mol. Biol.307, 541 (2001).
39
W. Yang, T. A. Steitz, Structure3, 131 (1995).
40
D. S. Schwarz, Y. Tomari, P. D. Zamore, Curr. Biol.14, 787 (2004).
41
R. F. Ketting, T. H. Haverkamp, H. G. van Luenen, R. H. Plasterk, Cell99, 133 (1999).
42
T. Sijen, R. H. Plasterk, Nature426, 310 (2003).
43
E. Sarot, G. Payen-Groschene, A. Bucheton, A. Pelisson, Genetics166, 1313 (2004).
44
Materials and methods are available as supporting material on Science Online.
45
The authors thank members of the Hannon lab for helpful discussions, Alea Mills for advice on ES cell work and for providing the library of targeting constructs, Sang Yong Kim for generating chimeras, Kathryn Anderson for insightful discussions and advice, and Phil Sharp for providing the CXCR4 constructs. M.C. is supported by the U.S. Army Breast Cancer Research Program, F.V.R. by the Jane Coffin Childs Memorial Fund, and J.S. by a Bristol Myers Squibb predoctoral fellowship. S.M.H. is a General Motors Cancer Research Foundation Scholar. This work was supported in part by grants from NIH (L.J. and G.J.H.).
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Science
Volume 305 | Issue 5689
3 September 2004
3 September 2004
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American Association for the Advancement of Science.
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Received: 8 July 2004
Accepted: 19 July 2004
Published in print: 3 September 2004
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