PUBLICATION

Translational inhibition by deadenylation-independent mechanisms is central to microRNA-mediated silencing in zebrafish

Authors
Mishima, Y., Fukao, A., Kishimoto, T., Sakamoto, H., Fujiwara, T., and Inoue, K.
ID
ZDB-PUB-120111-40
Date
2012
Source
Proceedings of the National Academy of Sciences of the United States of America   109(4): 1104-1109 (Journal)
Registered Authors
Inoue, Kunio, Kishimoto, Tomoyoshi, Mishima, Yuichiro
Keywords
none
MeSH Terms
  • Blotting, Western
  • Luciferases
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism
  • Autoantigens/genetics*
  • Autoantigens/metabolism
  • Zebrafish/embryology*
  • Zebrafish/metabolism
  • MicroRNAs/metabolism
  • MicroRNAs/physiology*
  • Green Fluorescent Proteins/metabolism
  • RNA Stability/physiology*
  • Gene Silencing/physiology*
  • Real-Time Polymerase Chain Reaction
  • In Situ Hybridization
  • Amino Acid Motifs/genetics
  • Mutagenesis, Site-Directed
  • Animals
  • Models, Biological*
  • Gene Expression Regulation, Developmental/physiology*
(all 20)
PubMed
22232654 Full text @ Proc. Natl. Acad. Sci. USA
Abstract
MicroRNA (miRNA) is a class of small noncoding RNA approximately 22 nt in length. Animal miRNA silences complementary mRNAs via translational inhibition, deadenylation, and mRNA degradation. However, the underlying molecular mechanisms remain unclear. A key question is whether these three outputs are independently induced by miRNA through distinct mechanisms or sequentially induced within a single molecular pathway. Here, we successfully dissected these intricate outputs of miRNA-mediated repression using zebrafish embryos as a model system. Our results indicate that translational inhibition and deadenylation are independent outputs mediated by distinct domains of TNRC6A, which is an effector protein in the miRNA pathway. Translational inhibition by TNRC6A is divided into two mechanisms: PAM2 motif-mediated interference of poly(A)-binding protein (PABP), and inhibition of 52 cap- and poly(A) tail-independent step(s) by a previously undescribed P-GL motif. Consistent with these observations, we show that, in zebrafish embryos, miRNA inhibits translation of the target mRNA in a deadenylation- and PABP-independent manner at early time points. These results indicate that miRNA exerts multiple posttranscriptional outputs via physically and functionally independent mechanisms and that direct translational inhibition is central to miRNA-mediated repression.
Genes / Markers
Figures
Figure Gallery (11 images) / 2
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Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
kb2TgTransgenic Insertion
    kb3TgTransgenic Insertion
      ya101TgTransgenic Insertion
        1 - 3 of 3
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        Human Disease / Model
        No data available
        Sequence Targeting Reagents
        Target Reagent Reagent Type
        mir1-1MO1-mir1-1,mir1-2MRPHLNO
        mir1-2MO1-mir1-1,mir1-2MRPHLNO
        mir206-1MO1-mir206-1MRPHLNO
        pabpc1aMO1-pabpc1aMRPHLNO
        1 - 4 of 4
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        Fish
        Antibodies
        Name Type Antigen Genes Isotypes Host Organism
        Ab1-mycmonoclonal
          IgG1Mouse
          Ab1-pabpcpolyclonal
            Rabbit
            1 - 2 of 2
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            Orthology
            No data available
            Engineered Foreign Genes
            Marker Marker Type Name
            GFPEFGGFP
            1 - 1 of 1
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            Mapping
            No data available