Regulation of the Elongation Phase of Protein Synthesis Enhances Translation Accuracy and Modulates Lifespan

Xie, Jianling; Alves, Viviane S.; Haar, Tobias von der; O’Keefe, Louise V.; Lenchine, Roman V.; Jensen, Kirk B.; Li, Rui; Coldwell, Mark J.; Wang, Xuemin; Proud, Christopher G.

doi:10.1016/j.cub.2019.01.029

Cited by 66 publications

(69 citation statements)

References 62 publications

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“…In contrast, the presence or absence of eEF2k did not affect the Fluc levels observed with the vector which has AUG as the start of the Fluc cistron ( Fig. S4A) which is in line with our earlier data [28].…”

Section: Eef2k Enhances Translation Of the Pd-l1 Mrna By Bypassing Trsupporting

confidence: 92%

“…The uORF partially overlaps with and inhibits the translation of the main ORF encoding PD-L1 [26]. We previously showed that activation of eEF2K helps reduce initiation at non-canonical start sites, i.e., CUG or GUG [28]. Impaired elongation slows down ribosomes within the main ORF and thus allows more time for selection of the optimal choice -an AUG start codon.…”

Section: Discussionmentioning

confidence: 99%

“…S2B). [Note that, upon activation, eEF2K is sometimes degraded (via the proteasome [19,28]). Similarly, PD-L1 levels were also reduced in eEF2K-null MDA-MB-231 cells ( Fig.…”

Section: Eef2k Promotes the Translation Of Pd-l1mentioning

confidence: 99%

“…(In contrast, the mouse Pd-l1 mRNA contains three possible uORFs starting with either an AUG or a non-canonical CUG (uCUG) [26]). We recently reported that active eEF2K, and thus slower elongation, favors the selection of the canonical AUG as a start codon by augmenting the ability of ribosomes to distinguish between AUG and near-cognate start codons [28].…”

Section: Eef2k Enhances Translation Of the Pd-l1 Mrna By Bypassing Trmentioning

confidence: 99%

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eEF2 kinase enhances the expression of PD-L1 by promoting the translation of its mRNA

Xie

Jin

et al. 2020

Preprint

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Emerging advances in cancer therapy have transformed the landscape from conventional therapies towards cancer immunotherapy regimens. Recent discoveries have resulted in the development of clinical immune checkpoint inhibitors that are ‘game-changers’ for cancer immunotherapy. Here we show that eEF2K, an atypical protein kinase that inhibits the elongation stage of protein synthesis, actually promotes the synthesis of PD-L1, an immune checkpoint protein which helps cancer cells to escape from immunosurveillance. Ablation of eEF2K in prostate and lung cancer cells markedly reduced the expression levels of the PD-L1 protein. We show that eEF2K promotes the association of PD-L1 mRNAs with translationally active polyribosomes and that translation of the PD-L1 mRNA is regulated by a uORF (upstream open reading-frame) within its 5’-UTR (5’-untranslated region) which starts with a non-canonical CUG codon. This inhibitory effect is attenuated by eEF2K thereby allowing higher levels of translation of the PD-L1 coding region and enhanced expression of the PD-L1 protein. Moreover, eEF2K-depleted cancer cells are more vulnerable to immune attack by natural killer cells. Therefore, control of translation elongation can modulate the translation of this specific mRNA, one which contains an uORF that starts with CUG, and perhaps others that contain a similar feature. Taken together, our data reveal that eEF2K regulates PD-L1 expression at the level of the translation of its mRNA by virtue of a uORF in its 5’-region. This, and other roles of eEF2K in cancer cell biology (e.g., in cell survival and migration), may be exploited for the design of future therapeutic strategies.

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Section: Eef2k Enhances Translation Of the Pd-l1 Mrna By Bypassing Trsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

“…S2B). [Note that, upon activation, eEF2K is sometimes degraded (via the proteasome [19,28]). Similarly, PD-L1 levels were also reduced in eEF2K-null MDA-MB-231 cells ( Fig.…”

Section: Eef2k Promotes the Translation Of Pd-l1mentioning

confidence: 99%

Section: Eef2k Enhances Translation Of the Pd-l1 Mrna By Bypassing Trmentioning

confidence: 99%

See 2 more Smart Citations

eEF2 kinase enhances the expression of PD-L1 by promoting the translation of its mRNA

Xie

Jin

et al. 2020

Preprint

Self Cite

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“…Cell lysates were heated at 95°C for 5 min in Laemmli Sample Buffer and then subjected to sodium-dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE [29];) followed by electrophoretic transfer to 0.45 µm nitrocellulose membranes (Bio-Rad; 1620115). Membranes were blocked in PBS-0.05% Tween20 (PBST) containing 5% (w/ v) skim milk powder for 60 min at room temperature.…”

Section: Immunoblot Analysesmentioning

confidence: 99%

Identification of DNA response elements regulating expression of CCAAT/enhancer-binding protein (C/EBP) β and δ and MAP kinase-interacting kinases during early adipogenesis

et al. 2020

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Given the high and increasing prevalence of obesity and associated disorders, such as type-2 diabetes, it is important to understand the mechanisms that regulate lipid storage and the differentiation of fat cells, a process termed adipogenesis. Using the well-established mouse 3T3-L1 in vitro model of adipogenesis, we refine how the induction of two key adipogenic transcription factors, CCAAT/enhancer-binding proteins (C/EBPs) β and δ are regulated during early adipogenesis. We identify, in the gene promoters of Cebpb and Cebpd, the DNA response elements responsible for binding transcription factors that are activated by cAMP or glucocorticoids. We also show that mitogen-activated protein kinase (MAPK)-interacting kinase 2 (MNK2; Mknk2), which plays a distinct role in diet-induced obesity, is induced during early adipogenesis and identify the functional DNA response elements responsible for regulating its expression. Mknk2 expression is maintained in differentiated 3T3-L1 adipocytes and is expressed at high levels across a range of mouse adipose tissue depots. Together, these new insights help to clarify the transcriptional programme of early adipogenesis and identify Mknk2 as one of potentially many genes up-regulated during adipogenesis.

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The mTOR–lysosome axis at the centre of ageing

et al. 2021

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Age‐related diseases represent some of the largest unmet clinical needs of our time. While treatment of specific disease‐related signs has had some success (for example, the effect of statin drugs on slowing progression of atherosclerosis), slowing biological ageing itself represents a target that could significantly increase health span and reduce the prevalence of multiple age‐related diseases. Mechanistic target of rapamycin complex 1 (mTORC1) is known to control fundamental processes in ageing: inhibiting this signalling complex slows biological ageing, reduces age‐related disease pathology and increases lifespan in model organisms. How mTORC1 inhibition achieves this is still subject to ongoing research. However, one mechanism by which mTORC1 inhibition is thought to slow ageing is by activating the autophagy–lysosome pathway. In this review, we examine the special bidirectional relationship between mTORC1 and the lysosome. In cells, mTORC1 is located on lysosomes. From this advantageous position, it directly controls the autophagy–lysosome pathway. However, the lysosome also controls mTORC1 activity in numerous ways, creating a special two‐way relationship. We then explore specific examples of how inhibition of mTORC1 and activation of the autophagy–lysosome pathway slow the molecular hallmarks of ageing. This body of literature demonstrates that the autophagy–lysosome pathway represents an excellent target for treatments that seek to slow biological ageing and increase health span in humans.

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Regulation of the Elongation Phase of Protein Synthesis Enhances Translation Accuracy and Modulates Lifespan

Cited by 66 publications

References 62 publications

eEF2 kinase enhances the expression of PD-L1 by promoting the translation of its mRNA

eEF2 kinase enhances the expression of PD-L1 by promoting the translation of its mRNA

Identification of DNA response elements regulating expression of CCAAT/enhancer-binding protein (C/EBP) β and δ and MAP kinase-interacting kinases during early adipogenesis

The mTOR–lysosome axis at the centre of ageing

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