A Slicer-Mediated Mechanism for Repeat-Associated siRNA 5' End Formation in Drosophila

American Association for the Advancement of Science (AAAS) - Tập 315 Số 5818 - Trang 1587-1590 - 2007
Lalith Gunawardane1, Kuniaki Saito1, Kazumichi M. Nishida1, Keita Miyoshi1, Yoshinori Kawamura1, Tomoko Nagami1, Haruhiko Siomi1, Mikiko C. Siomi1
1Institute for Genome Research, University of Tokushima, Tokushima 770-8503, Japan

Tóm tắt

In Drosophila , repeat-associated small interfering RNAs (rasiRNAs) are produced in the germ line by a Dicer-independent pathway and function through the PIWI subfamily of Argonautes to ensure silencing of retrotransposons. We sequenced small RNAs associated with the PIWI subfamily member AGO3. Although other members of PIWI, Aubergine (Aub) and Piwi, associated with rasiRNAs derived mainly from the antisense strand of retrotransposons, AGO3-associated rasiRNAs arose mainly from the sense strand. Aub- and Piwi-associated rasiRNAs showed a strong preference for uracil at their 5′ ends, and AGO3-associated rasiRNAs showed a strong preference for adenine at nucleotide 10. Comparisons between AGO3- and Aub-associated rasiRNAs revealed pairs of rasiRNAs showing complementarities in their first 10 nucleotides. Aub and AGO3 exhibited Slicer activity in vitro. These data support a model in which formation of a 5′ terminus within rasiRNA precursors is guided by rasiRNAs originating from transcripts of the other strand in concert with the Slicer activity of PIWI.

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We thank T. Suzuki and V. V. Vagin for technical advice; T. Mori M. Shinomiya and A. Kinouchi for assistance with fly husbandry and dissection; and members of the Siomi laboratory for discussions and comments on the manuscript. Supported by the Ministry of Education Culture Sports Science and Technology of Japan (M.C.S. H.S.) and its 21st Century Centers of Excellence Program (L.S.G. K.S.) the Japan Society for the Promotion of Science (JSPS) (M.C.S. and H.S.) a JSPS research fellowship (K.M.) and the New Energy and Industrial Technology Development Organization (M.C.S).

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American Association for the Advancement of Science (AAAS)

Tập 315 Số 5818

1587-1590

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