PUBLICATION

Epitranscriptomics m6 A analyses reveal distinct m6 A marks under tumor necrosis factor α (TNF-α)-induced apoptotic conditions in HeLa cells

Authors
Akçaöz-Alasar, A., Tüncel, Ö., Sa?lam, B., Gazalo?lu, Y., Atbinek, M., Cagiral, U., Iscan, E., Ozhan, G., Akgül, B.
ID
ZDB-PUB-240105-35
Date
2024
Source
Journal of Cellular Physiology   239(4): e31176 (Journal)
Registered Authors
Özhan, Günes
Keywords
Hela, RNA modification, TNF-alpha, apoptosis, epitranscriptomics, m6A
MeSH Terms
  • Apoptosis/genetics
  • Humans
  • HeLa Cells
  • Tumor Necrosis Factor-alpha*/metabolism
  • Tumor Necrosis Factor-alpha*/pharmacology
  • Animals
  • RNA
  • Methyltransferases/genetics
  • Methyltransferases/metabolism
  • Heat-Shock Proteins/metabolism
  • Zebrafish*/genetics
  • Zebrafish*/metabolism
  • Carrier Proteins/metabolism
  • Gene Expression Regulation
PubMed
38179601 Full text @ J. Cell. Physiol.
Abstract
Tumor necrosis factor-α (TNF-α) is a ligand that induces both intrinsic and extrinsic apoptotic pathways in HeLa cells by modulating complex gene regulatory mechanisms. However, the full spectrum of TNF-α-modulated epitranscriptomic m6 A marks is unknown. We employed a genomewide approach to examine the extent of m6 A RNA modifications under TNF-α-modulated apoptotic conditions in HeLa cells. miCLIP-seq analyses revealed a plethora of m6 A marks on 632 target mRNAs with an enrichment on 99 mRNAs associated with apoptosis. Interestingly, the m6 A RNA modification patterns were quite different under cisplatin- and TNF-α-mediated apoptotic conditions. We then examined the abundance and translational efficiencies of several mRNAs under METTL3 knockdown and/or TNF-α treatment conditions. Our analyses showed changes in the translational efficiency of TP53INP1 mRNA based on the polysome profile analyses. Additionally, TP53INP1 protein amount was modulated by METTL3 knockdown upon TNF-α treatment but not CP treatment, suggesting the existence of a pathway-specific METTL3-TP53INP1 axis. Congruently, METLL3 knockdown sensitized HeLa cells to TNF-α-mediated apoptosis, which was also validated in a zebrafish larval xenograft model. These results suggest that apoptotic pathway-specific m6 A methylation marks exist in cells and TNF-α-METTL3-TP53INP1 axis modulates TNF-α-mediated apoptosis in HeLa cells.
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Mutations / Transgenics
Human Disease / Model
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Mapping