3' end formation of PIWI-interacting RNAs in vitro.
Mol Cell
; 43(6): 1015-22, 2011 Sep 16.
Article
en En
| MEDLINE
| ID: mdl-21925389
ABSTRACT
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 Mg(2+)-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.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Bombyx
/
Procesamiento de Término de ARN 3'
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ARN Interferente Pequeño
Límite:
Animals
Idioma:
En
Año:
2011
Tipo del documento:
Article