m6A-modified cenRNA stabilizes CENPA to ensure centromere integrity in cancer cells.

m6A modification is best known for its critical role in controlling multiple post-transcriptional processes of the mRNAs. Here, we discovered elevated levels of m6A modification on centromeric RNA (cenRNA) in cancerous cells compared with non-cancerous cells. We then identified CENPA, an H3 variant, as an m6A reader of cenRNA. CENPA is localized at centromeres and is essential in preserving centromere integrity and function during mitosis. The m6A-modified cenRNA stabilizes centromeric localization of CENPA in cancer cells during the S phase of the cell cycle. Mutations of CENPA at the Leu61 and the Arg63 or removal of cenRNA m6A modification lead to loss of centromere-bound CENPA during S phase. This in turn results in compromised centromere integrity and abnormal chromosome separation and hinders cancer cell proliferation and tumor growth. Our findings unveil an m6A reading mechanism by CENPA that epigenetically governs centromere integrity in cancer cells, providing potential targets for cancer therapy. [Display omitted] • m6A modifications on centromeric RNA (cenRNA) are elevated in cancer cells • CENPA binding to m6A-cenRNA ensures its centromere association during S phase • The CENPA residues Leu61 and Arg63 mediate its interaction with m6A-cenRNA • Disruption of the CENPA-m6A-cenRNA interaction impairs cancer cell centromere integrity CENPA is an m6A reader with specificity for m6A-modified centromeric RNA. The CENPA-centromeric RNA interaction ensures centromere integrity and proliferation in cancer cells. [ABSTRACT FROM AUTHOR]