期刊
NATURE STRUCTURAL & MOLECULAR BIOLOGY
卷 18, 期 2, 页码 237-U309出版社
NATURE RESEARCH
DOI: 10.1038/nsmb.1991
关键词
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资金
- Leukemia and Lymphoma Society
- David H. Koch graduate fellowship
- US National Institutes of Health [R01-GM34277, R01-CA133404]
- National Cancer Institute [P01-CA42063, P30-CA14051]
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 Dicer(-/-) 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 similar to 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.
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