A slicer-mediated mechanism for repeat-associated siRNA 5′ end formation in Drosophila

Lalith S. Gunawardane, Kuniaki Saito, Kazumichi M. Nishida, Keita Miyoshi, Yoshinori Kawamura, Tomoko Nagami, Haruhiko Siomi, Mikiko C. Siomi

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Abstract

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 AG03. 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.

Original languageEnglish
Pages (from-to)1587-1590
Number of pages4
JournalScience
Volume315
Issue number5818
DOIs
Publication statusPublished - 2007 Mar 16

ASJC Scopus subject areas

  • General

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    Gunawardane, L. S., Saito, K., Nishida, K. M., Miyoshi, K., Kawamura, Y., Nagami, T., Siomi, H., & Siomi, M. C. (2007). A slicer-mediated mechanism for repeat-associated siRNA 5′ end formation in Drosophila. Science, 315(5818), 1587-1590. https://doi.org/10.1126/science.1140494