Genome-wide identification of Ago2 binding sites from mouse embryonic stem cells with and without mature microRNAs.
Nat Struct Mol Biol
; 18(2): 237-44, 2011 Feb.
Article
en En
| MEDLINE
| ID: mdl-21258322
ABSTRACT
MicroRNAs (miRNAs) are 19-22-nucleotide noncoding RNAs that post-transcriptionally regulate mRNA targets. We have identified endogenous miRNA binding sites in mouse embryonic stem cells (mESCs), by performing photo-cross-linking immunoprecipitation using antibodies to Argonaute (Ago2) followed by deep sequencing of RNAs (CLIP-seq). We also performed CLIP-seq in Dicerâ»/â» mESCs that lack mature miRNAs, allowing us to define whether the association of Ago2 with the identified sites was miRNA dependent. A significantly enriched motif, GCACUU, was identified only in wild-type mESCs in 3' untranslated and coding regions. This motif matches the seed of a miRNA family that constitutes ~68% of the mESC miRNA population. Unexpectedly, a G-rich motif was enriched in sequences cross-linked to Ago2 in both the presence and absence of miRNAs. Expression analysis and reporter assays confirmed that the seed-related motif confers miRNA-directed regulation on host mRNAs and that the G-rich motif can modulate this regulation.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Factor 2 Eucariótico de Iniciación
/
Regulación de la Expresión Génica
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MicroARNs
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Células Madre Embrionarias
Tipo de estudio:
Diagnostic_studies
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Prognostic_studies
Límite:
Animals
Idioma:
En
Revista:
Nat Struct Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2011
Tipo del documento:
Article
País de afiliación:
Estados Unidos