Abstract
During the integrated stress response (ISR), global translation initiation is attenuated; however, noncanonical mechanisms allow for the continued translation of specific transcripts. Eukaryotic initiation factor 5B (eIF5B) has been shown to play a critical role in canonical translation as well as in noncanonical mechanisms involving internal ribosome entry site (IRES) and upstream open reading frame (uORF) elements. The uORF-mediated translation regulation of activating transcription factor 4 (ATF4) mRNA plays a pivotal role in the cellular ISR. Our recent study confirmed that eIF5B depletion removes uORF2-mediated repression of ATF4 translation, which results in the upregulation of growth arrest and DNA damage-inducible protein 34 (GADD34) transcription. Accordingly, we hypothesized that eIF5B depletion may reprogram the transcriptome profile of the cell. Here, we employed genome-wide transcriptional analysis on eIF5B-depleted cells. Further, we validate the up- and downregulation of several transcripts from our RNA-seq data using RT-qPCR. We identified upregulated pathways including cellular response to endoplasmic reticulum (ER) stress, and mucin-type O-glycan biosynthesis, as well as downregulated pathways of transcriptional misregulation in cancer and T cell receptor signaling. We also confirm that depletion of eIF5B leads to activation of the c-Jun N-terminal kinase (JNK) arm of the mitogen-activated protein kinase (MAPK) pathway. This data suggests that depletion of eIF5B reprograms the cellular transcriptome and influences critical cellular processes such as ER stress and ISR.
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Funding
K.R.B was supported by NSERC-CGS and Queen Elizabeth II fellowships. K.V.D. was supported by Queen Elizabeth II and Alberta Innovates Tech Future graduate fellowships. K.P. and J.A.R, undergraduate and postdoctoral fellow, respectively, were partially supported by NSERC-CREATE grant (510937-2018). This work was funded by a Natural Sciences and Engineering Research Council of Canada-Discovery Grant (RGPIN-2017-05463), the Canada Foundation for Innovation-John R. Evans Leaders Fund (35017), the Campus Alberta Innovates Program, and the Alberta Ministry of Economic Development and Trade.
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Conceptualization, K.R.B. and N.T.; transcriptome data analysis, S.I., K.V.D., and A.Z.; investigation, K.R.B., J.A.R., K.V.D., K.T., and K.P.; resources, N.T., I.K., and A.Z.; writing and editing, K.R.B., J.A.R., and N.T.; funding acquisition, N.T.
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Bressler, K.R., Ross, J.A., Ilnytskyy, S. et al. Depletion of eukaryotic initiation factor 5B (eIF5B) reprograms the cellular transcriptome and leads to activation of endoplasmic reticulum (ER) stress and c-Jun N-terminal kinase (JNK). Cell Stress and Chaperones 26, 253–264 (2021). https://doi.org/10.1007/s12192-020-01174-1
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DOI: https://doi.org/10.1007/s12192-020-01174-1