期刊
MOLECULAR CELL
卷 43, 期 6, 页码 1015-1022出版社
CELL PRESS
DOI: 10.1016/j.molcel.2011.07.029
关键词
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资金
- Functional machinery for non-coding RNAs
- Innovative Areas (Functional machinery for non-coding RNAs)
- Grants-in-Aid for Scientific Research [22115502] Funding Source: KAKEN
PIWI-interacting RNAs (piRNAs) are 23-30 nucleotides small RNAs that act with PIWI proteins to silence transposon activity in animal gonads. In contrast to microRNAs and small interfering RNAs, the biogenesis of piRNAs, including how 3' ends are formed, remains largely unknown. Here, by using lysate from BmN4, a silkworm ovary-derived cell line, we have developed a cell-free system that recapitulates key steps of piRNA biogenesis: loading of long single-stranded precursor RNAs into PIWI proteins with 5'-nucleotide bias, followed by Mg2+-dependent 3' to 5' exonucleolytic trimming and 2'-O-methylation at 3' ends. Importantly, 3' end methylation is tightly coupled with trimming and yet is not a prerequisite for determining the mature piRNA length. Our system provides a biochemical framework for dissecting piRNA biogenesis.
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