Work

Quantification of Propargylated RNA Nucleosides After Metabolic Labeling Via the Methylation Pathway

Year: 2025

Type: article

Abstract: RNA modifications are involved in numerous biological processes and vary in different cell types. Methylation is the most widespread type of RNA modification and occurs via S ‐adenosyl‐L‐methionine (SAM). We recently developed a metabolic labeling approach based on intracellular formation of a clickable SAM analog (SeAdoYn) and demonstrated its use in mapping methyltransferase (MTase) target sites in mRNA from HeLa cells. Here we investigate how metabolic labeling via the clickable SAM analog modifies four different nucleosides in RNA of HEK293T in comparison to HeLa cells. We find that HEK293T cells retain higher cell viability upon feeding the clickable metabolic SAM precursor. In poly(A) + RNA we find high A prop /A levels (0.04 %) and in total RNA (but not poly(A) + RNA) we detect prop 3 C, which had not been detected previously in HeLa cells. We discuss the findings in the context of data from the literature with respect to mRNA half‐lives in cancer and non‐cancer cell lines and suggest that CMTr2 is most likely responsible for the high A prop level in poly(A) + RNA.

Source: ChemBioChem

DOI: 10.1002/cbic.202400986

Authors: Nadine A. Kueck, Sabine Hüwel, Arne Hoffmann, Andrea Rentmeister

Institutions: University of Münster, Ludwig-Maximilians-Universität München

Language: English

Topic: RNA modifications and cancer

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: hybrid

Estimated APC paid: $4,430