TY - JOUR
T1 - Crystal structure of Drosophila Piwi
AU - Yamaguchi, Sonomi
AU - Oe, Akira
AU - Nishida, Kazumichi M.
AU - Yamashita, Keitaro
AU - Kajiya, Asako
AU - Hirano, Seiichi
AU - Matsumoto, Naoki
AU - Dohmae, Naoshi
AU - Ishitani, Ryuichiro
AU - Saito, Kuniaki
AU - Siomi, Haruhiko
AU - Nishimasu, Hiroshi
AU - Siomi, Mikiko C.
AU - Nureki, Osamu
N1 - Funding Information:
We thank Dr. Masahiro Fukuda, Dr. Wataru Shihoya, and the beamline scientists at SPring-8 BL32XU for assistance with data collection, Dr. Takanori Nakane for assistance with structure determination, and Atsuhiro Tomita for assistance with the ITC experiments. This work was supported by JSPS KAKENHI Grant Numbers 17H05592 (H.N.), 18H02384 (H.N.), and 19H05466 (M.C.S.).
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - PIWI-clade Argonaute proteins associate with PIWI-interacting RNAs (piRNAs), and silence transposons in animal gonads. Here, we report the crystal structure of the Drosophila PIWI-clade Argonaute Piwi in complex with endogenous piRNAs, at 2.9 Å resolution. A structural comparison of Piwi with other Argonautes highlights the PIWI-specific structural features, such as the overall domain arrangement and metal-dependent piRNA recognition. Our structural and biochemical data reveal that, unlike other Argonautes including silkworm Siwi, Piwi has a non-canonical DVDK tetrad and lacks the RNA-guided RNA cleaving slicer activity. Furthermore, we find that the Piwi mutant with the canonical DEDH catalytic tetrad exhibits the slicer activity and readily dissociates from less complementary RNA targets after the slicer-mediated cleavage, suggesting that the slicer activity could compromise the Piwi-mediated co-transcriptional silencing. We thus propose that Piwi lost the slicer activity during evolution to serve as an RNA-guided RNA-binding platform, thereby ensuring faithful co-transcriptional silencing of transposons.
AB - PIWI-clade Argonaute proteins associate with PIWI-interacting RNAs (piRNAs), and silence transposons in animal gonads. Here, we report the crystal structure of the Drosophila PIWI-clade Argonaute Piwi in complex with endogenous piRNAs, at 2.9 Å resolution. A structural comparison of Piwi with other Argonautes highlights the PIWI-specific structural features, such as the overall domain arrangement and metal-dependent piRNA recognition. Our structural and biochemical data reveal that, unlike other Argonautes including silkworm Siwi, Piwi has a non-canonical DVDK tetrad and lacks the RNA-guided RNA cleaving slicer activity. Furthermore, we find that the Piwi mutant with the canonical DEDH catalytic tetrad exhibits the slicer activity and readily dissociates from less complementary RNA targets after the slicer-mediated cleavage, suggesting that the slicer activity could compromise the Piwi-mediated co-transcriptional silencing. We thus propose that Piwi lost the slicer activity during evolution to serve as an RNA-guided RNA-binding platform, thereby ensuring faithful co-transcriptional silencing of transposons.
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U2 - 10.1038/s41467-020-14687-1
DO - 10.1038/s41467-020-14687-1
M3 - Article
C2 - 32051406
AN - SCOPUS:85079337901
SN - 2041-1723
VL - 11
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 858
ER -