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Title: 28S rRNA is inducibly pseudouridylated by the mTOR pathway translational control in CHO cell cultures
Authors: Courtes, F.C.
Gu, C.
Wong, N.S.C.
Dedon, P.C.
Yap, M.G.S. 
Lee, D.-Y. 
Keywords: CHO cells culture
Inducible pseudouridylation
MTOR pathway
SnoRNA U19
Issue Date: 20-Mar-2014
Citation: Courtes, F.C., Gu, C., Wong, N.S.C., Dedon, P.C., Yap, M.G.S., Lee, D.-Y. (2014-03-20). 28S rRNA is inducibly pseudouridylated by the mTOR pathway translational control in CHO cell cultures. Journal of Biotechnology 174 (1) : 16-21. ScholarBank@NUS Repository.
Abstract: The mTOR pathway is a conserved master regulator of translational activity that influences the fate of industrially relevant CHO cell cultures, yet its molecular mechanisms remain unclear. Interestingly, rapamycin specific inhibition of the mTOR pathway in CHO cells was found to down-regulate the small nucleolar RNA U19 (snoRNA U19) by 2-fold via translatome profiling. snoRNA U19 guides the two most conserved pseudouridylation modifications on 28S ribosomal RNA (rRNA) that are important for the biogenesis and proper function of ribosomes. In order to further understand the role of snoRNA U19 as a potential player in the mTOR pathway, we measured 28S rRNA pseudouridylation upon rapamycin treatments and/or snoRNA U19 overexpression conditions, thereby characterizing the subsequent effects on ribosome efficiency and global translation by polysome profiling. We showed that 28S rRNA pseudouridylation was increased by rapamycin treatment and/or overexpression of snoRNA U19, but only the latter condition improved ribosome efficiency toward higher global translation, thus implying that the mTOR pathway induces pseudouridylation at different sites along the 28S rRNA possibly with either positive or negative effects on the cellular phenotype. This discovery of snoRNA U19 as a new downstream effector of the mTOR pathway suggests that cell engineering of snoRNAs can be used to regulate translation and improve cellular growth in CHO cell cultures in the future. © 2014 Elsevier B.V.
Source Title: Journal of Biotechnology
ISSN: 01681656
DOI: 10.1016/j.jbiotec.2014.01.024
Appears in Collections:Staff Publications

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