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RNA Biol. 2013 Jul;10(7):1146-59. doi: 10.4161/rna.25194. Epub 2013 May 29.

Identification of LIN28B-bound mRNAs reveals features of target recognition and regulation.

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Systems Biology of Gene Regulatory Elements; Max-Delbrück-Center for Molecular Medicine; Berlin, Germany.


The conserved human LIN28 RNA-binding proteins function in development, maintenance of pluripotency and oncogenesis. We used PAR-CLIP and a newly developed variant of this method, iDo-PAR-CLIP, to identify LIN28B targets as well as sites bound by the individual RNA-binding domains of LIN28B in the human transcriptome at nucleotide resolution. The position of target binding sites reflected the known structural relative orientation of individual LIN28B-binding domains, validating iDo-PAR-CLIP. Our data suggest that LIN28B directly interacts with most expressed mRNAs and members of the let-7 microRNA family. The Lin28-binding motif detected in pre-let-7 was enriched in mRNA sequences bound by LIN28B. Upon LIN28B knockdown, cell proliferation and the cell cycle were strongly impaired. Quantitative shotgun proteomics of LIN28B depleted cells revealed significant reduction of protein synthesis from its RNA targets. Computational analyses provided evidence that the strength of protein synthesis reduction correlated with the location of LIN28B binding sites within target transcripts.


CLIP; RNA-binding protein; post-transcriptional regulation; stem cell

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