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Nucleic Acids Res ; 46(22): 12109-12125, 2018 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-30295819

RESUMO

Post-transcriptional gene regulation controls the amount of protein produced from a specific mRNA by altering both its decay and translation rates. Such regulation is primarily achieved by the interaction of trans-acting factors with cis-regulatory elements in the untranslated regions (UTRs) of mRNAs. These interactions are guided either by sequence- or structure-based recognition. Similar to sequence conservation, the evolutionary conservation of a UTR's structure thus reflects its functional importance. We used such structural conservation to identify previously unknown cis-regulatory elements. Using the RNA folding program Dynalign, we scanned all UTRs of humans and mice for conserved structures. Characterizing a subset of putative conserved structures revealed a binding site of the RNA-binding protein Roquin. Detailed functional characterization in vivo enabled us to redefine the binding preferences of Roquin and identify new target genes. Many of these new targets are unrelated to the established role of Roquin in inflammation and immune responses and thus highlight additional, unstudied cellular functions of this important repressor. Moreover, the expression of several Roquin targets is highly cell-type-specific. In consequence, these targets are difficult to detect using methods dependent on mRNA abundance, yet easily detectable with our unbiased strategy.


Assuntos
Biologia Computacional/métodos , Regulação da Expressão Gênica , Dobramento de RNA , Proteínas de Ligação a RNA/química , Ubiquitina-Proteína Ligases/química , Regiões 3' não Traduzidas , Animais , Sítios de Ligação , Linhagem Celular , Simulação por Computador , Análise Mutacional de DNA , Células HEK293 , Células HeLa , Humanos , Camundongos , Conformação de Ácido Nucleico , Nucleotídeos/genética , Ligação Proteica , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/genética , Transcrição Gênica , Ubiquitina-Proteína Ligases/genética
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