Transcriptomics

Dataset Information

355

PARN mediates 3'-end trimming of Argonaute-cleaved precursor microRNAs


ABSTRACT: microRNAs (miRNAs) are typically generated as ~22-nucleotide double-stranded RNAs via processing of precursor hairpins by the RNase III enzyme Dicer, after which they are loaded into Argonaute (Ago) proteins to form RNA-induced silencing complex (RISC). However, the biogenesis of miR-451, an erythropoietic miRNA conserved in vertebrates, does not require Dicer processing. Instead, the short pre-miR-451 precursor hairpin is directly loaded into Ago, followed by cleavage of the 3' arm and trimming of the 3' end to the mature length by PARN. Here we show the in vivo activity of miR-430 Ago2-hairpin, a canonical microRNA engineered to fit the structure of miR-451 and hence become Ago2-dependent. Moreover, we test a modified miR-430 Ago2-haipin with 3x phoshorothioate bonds that impairs trimmng. Surprisingly, our data show that trimming of Ago-cleaved pre-miRNAs is not essential for target silencing, indicating that RISC is functional with miRNAs longer than 22-nucleotides. Rescue of MZdicer zebrafish mutant with the injection of trimmable and nontrimmable miR-430 Ago2 hairpins: Transcriptome of wild type, MZdicer mutant, and MZdicer mutant micoinjected with miR-430 duplex, miR-430 (Ago2-haripin), miR-430 (Ago2-haripin 3xPhosphorothioate)

ORGANISM(S): Danio rerio  

SUBMITTER: Yuriko Sakaguchi   Hukihide Tomari  Daniel Cifuentes  Natsuko Izumi  Antonio Giraldez  Mayuko Yoda  Tsutomu Suzuki 

PROVIDER: E-GEOD-51036 | ArrayExpress | 2013-11-13

SECONDARY ACCESSION(S): GSE51036SRP030117PRJNA219729

REPOSITORIES: GEO, ArrayExpress, ENA

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Publications

Poly(A)-specific ribonuclease mediates 3'-end trimming of Argonaute2-cleaved precursor microRNAs.

Yoda Mayuko M   Cifuentes Daniel D   Izumi Natsuko N   Sakaguchi Yuriko Y   Suzuki Tsutomu T   Giraldez Antonio J AJ   Tomari Yukihide Y  

Cell reports 20131024 3


MicroRNAs (miRNAs) are typically generated as ~22-nucleotide double-stranded RNAs via the processing of precursor hairpins by the ribonuclease III enzyme Dicer, after which they are loaded into Argonaute (Ago) proteins to form an RNA-induced silencing complex (RISC). However, the biogenesis of miR-451, an erythropoietic miRNA conserved in vertebrates, occurs independently of Dicer and instead requires cleavage of the 3' arm of the pre-miR-451 precursor hairpin by Ago2. The 3' end of the Ago2-cle  ...[more]

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