In Vivo RNAi-Mediated eIF3m Knockdown Affects Ribosome Biogenesis and Transcription but Has Limited Impact on mRNA-Specific Translation
Name
1-s2.0-S2162253119303622-main.pdf
Description
Published version
Size
2.36 MB
Format
Adobe PDF
Checksum (MD5)
6777e549405934c19348084d738b07c4
Author(s)
Smekalova, Elena M
Gerashchenko, Maxim V
O’Connor, Patrick BF
Whittaker, Charles A
Kauffman, Kevin J
Fefilova, Anna S
Zatsepin, Timofei S
Bogorad, Roman L
Baranov, Pavel V
Langer, Robert
Date Issued
2020
Journal
Molecular Therapy – Nucleic Acids
Publisher
Elsevier BV
Version
Final published version
Abstract
© 2019 Translation is an essential biological process, and dysregulation is associated with a range of diseases including ribosomopathies, diabetes, and cancer. Here, we examine translation dysregulation in vivo using RNAi to knock down the m-subunit of the translation initiation factor eIF3 in the mouse liver. Transcriptome sequencing, ribosome profiling, whole proteome, and phosphoproteome analyses show that eIF3m deficiency leads to the transcriptional response and changes in cellular translation that yield few detectable differences in the translation of particular mRNAs. The transcriptional response fell into two main categories: ribosome biogenesis (increased transcription of ribosomal proteins) and cell metabolism (alterations in lipid, amino acid, nucleic acid, and drug metabolism). Analysis of ribosome biogenesis reveals inhibition of rRNA processing, highlighting decoupling of rRNA synthesis and ribosomal protein gene transcription in response to eIF3m knockdown. Interestingly, a similar reduction in eIF3m protein levels is associated with induction of the mTOR pathway in vitro but not in vivo. Overall, this work highlights the utility of a RNAi-based in vivo approach for studying the regulation of mammalian translation in vivo.
MIT Department
Koch Institute for Integrative Cancer Research at MIT
Massachusetts Institute of Technology. Department of Chemical Engineering
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
10.1016/J.OMTN.2019.11.009
https://doi.org/10.1016/J.OMTN.2019.11.009
