WFRF:(Banijamali M)
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Efficient 3′-pairing renders microRNA targeting less sensitive to mRNA seed accessibility
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- Kosek, David M (author)
- Karolinska Institutet,Department of Cell and Molecular Biology, Karolinska Institute , Biomedicum 9B, Solnavägen 9, 17177 Stockholm , Sweden
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- Banijamali, Elnaz (author)
- Department of Medical Biochemistry and Biophysics, Karolinska Institute , Biomedicum 9B, Solnavägen 9, 17177 Stockholm , Sweden
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- Becker, Walter (author)
- Department of Medical Biochemistry and Biophysics, Karolinska Institute , Biomedicum 9B, Solnavägen 9, 17177 Stockholm , Sweden
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- (creator_code:org_t)
- Oxford University Press, 2023
- 2023
- English.
- In: Nucleic Acids Research. - : Oxford University Press. - 0305-1048 .- 1362-4962. ; 51:20, s. 11162-11177
- Journal article (peer-reviewed)
Abstract
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- MicroRNAs (miRNAs) are short RNAs that post-transcriptionally regulate gene expression by binding to specific sites in mRNAs. Site recognition is primarily mediated by the seed region (nucleotides g2–g8 in the miRNA), but pairing beyond the seed (3′-pairing) is important for some miRNA:target interactions. Here, we use SHAPE, luciferase reporter assays and transcriptomics analyses to study the combined effect of 3′-pairing and secondary structures in mRNAs on repression efficiency. Using the interaction between miR-34a and its SIRT1 binding site as a model, we provide structural and functional evidence that 3′-pairing can compensate for low seed-binding site accessibility, enabling repression of sites that would otherwise be ineffective. We show that miRNA 3′-pairing regions can productively base-pair with nucleotides far upstream of the seed-binding site and that both hairpins and unstructured bulges within the target site are tolerated. We use SHAPE to show that sequences that overcome inaccessible seed-binding sites by strong 3′-pairing adopt the predicted structures and corroborate the model using luciferase assays and high-throughput modelling of 8177 3′-UTR targets for six miRNAs. Finally, we demonstrate that PHB2, a target of miR-141, is an inaccessible target rescued by efficient 3′-pairing. We propose that these results could refine predictions of effective target sites.
Subject headings
- NATURVETENSKAP -- Biologi -- Biofysik (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biophysics (hsv//eng)
- NATURVETENSKAP -- Biologi -- Strukturbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Structural Biology (hsv//eng)
- NATURVETENSKAP -- Biologi -- Cellbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Cell Biology (hsv//eng)
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