5-Azacytidine incorporation into mRNAs disrupts translation and induces ribosome collisions
Abstract
5-Azacytidine (5-AzaC) is a cytidine analog chemotherapeutic widely used to treat myeloid malignancies. Although its activity is primarily attributed to DNA hypomethylation, the majority of 5-AzaC is incorporated into RNA, where its functional consequences are less understood. Here, we show that 5-AzaC treatment of cells leads to inhibition of protein synthesis. Ribo-seq, Disome-seq, and RNA-seq in treated cells exhibit a time-dependent C-to-G conversion in mRNAs. These conversion events are enriched within footprint positions corresponding to the A-site of monosomes or stalled leading ribosomes in a disome complex. Footprints are accumulated at sites with C-rich codons in the A-site, with particular emphasis on the second codon position. Consistent with disome-mediated signaling, 5-AzaC activates ZAK- and GCN2-dependent stress responses. Furthermore, loss of the ribosome-associated quality control factor, ZNF598, sensitizes cells to 5-AzaC. These findings suggest that translation disruption represents an additional layer of 5-AzaC’s mechanism of action, alongside its known DNA-mediated effects.
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BibTeX
@article{roberson20265azacytidine,
title = {{5-Azacytidine incorporation into mRNAs disrupts translation and induces ribosome collisions}},
author = {Alexis B. Roberson and James Marks and Ruby Pitts and Bavavarshini Tamilselvam and Grace D. Britt and Brian Grieb and William P. Tansey and Sezen Meydan},
journal = {Cell Reports},
year = {2026},
month = nov,
doi = {10.1016/j.celrep.2026.118247},
url = {https://linkinghub.elsevier.com/retrieve/pii/S2211124726013264},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - 5-Azacytidine incorporation into mRNAs disrupts translation and induces ribosome collisions
AU - Alexis B. Roberson
AU - James Marks
AU - Ruby Pitts
AU - Bavavarshini Tamilselvam
AU - Grace D. Britt
AU - Brian Grieb
AU - William P. Tansey
AU - Sezen Meydan
PY - 2026
DA - 2026/11/01
JO - Cell Reports
DO - 10.1016/j.celrep.2026.118247
UR - https://linkinghub.elsevier.com/retrieve/pii/S2211124726013264
AB - 5-Azacytidine (5-AzaC) is a cytidine analog chemotherapeutic widely used to treat myeloid malignancies. Although its activity is primarily attributed to DNA hypomethylation, the majority of 5-AzaC is incorporated into RNA, where its functional consequences are less understood. Here, we show that 5-AzaC treatment of cells leads to inhibition of protein synthesis. Ribo-seq, Disome-seq, and RNA-seq in treated cells exhibit a time-dependent C-to-G conversion in mRNAs. These conversion events are enriched within footprint positions corresponding to the A-site of monosomes or stalled leading ribosomes in a disome complex. Footprints are accumulated at sites with C-rich codons in the A-site, with particular emphasis on the second codon position. Consistent with disome-mediated signaling, 5-AzaC activates ZAK- and GCN2-dependent stress responses. Furthermore, loss of the ribosome-associated quality control factor, ZNF598, sensitizes cells to 5-AzaC. These findings suggest that translation disruption represents an additional layer of 5-AzaC’s mechanism of action, alongside its known DNA-mediated effects.
SN - 2211-1247
ER -