biostudies-literatureYan QFoundation for the National Institutes of HealthNational Natural Science Foundation of ChinaChina Postdoctoral Science FoundationNCI NIH HHSNatural Science Foundation of Jiangsu Province6327https://www.ebi.ac.uk/biostudies/studies/S-EPMC10564894biostudies-literatureUnknown14(1)N-acetyltransferase 10 (NAT10) is an N⁴-acetylcytidine (ac⁴C) writer that catalyzes RNA acetylation at cytidine N⁴ position on tRNAs, rRNAs and mRNAs. Recently, NAT10 and the associated ac⁴C have been reported to increase the stability of HIV-1 transcripts. Here, we show that NAT10 catalyzes ac⁴C addition to the polyadenylated nuclear RNA (PAN), a long non-coding RNA encoded by the oncogenic DNA virus Kaposi's sarcoma-associated herpesvirus (KSHV), triggering viral lytic reactivation from latency. Mutagenesis of ac⁴C sites in PAN RNA in the context of KSHV infection abolishes PAN ac⁴C modifications, downregulates the expression of viral lytic genes and reduces virion production. NAT10 knockdown or mutagenesis erasNature communicationsNAT10-dependent N⁴-acetylcytidine modification mediates PAN RNA stability, KSHV reactivation, and IFI16-related inflammasome activation.PMC10564894BK2018008482372243R01 CA213275822410678173006281761128003BK2018068181902639821722622019TQ01583180014832170151R01CA2132752019M6618898187164282072275Li WDing XGao SJYan QMa XLu CJia XWang ZZhou JfalseNAT10-dependent N⁴-acetylcytidine modification mediates PAN RNA stability, KSHV reactivation, and IFI16-related inflammasome activation.N-acetyltransferase 10 (NAT10) is an N⁴-acetylcytidine (ac⁴C) writer that catalyzes RNA acetylation at cytidine N⁴ position on tRNAs, rRNAs and mRNAs. Recently, NAT10 and the associated ac⁴C have been reported to increase the stability of HIV-1 transcripts. Here, we show that NAT10 catalyzes ac⁴C addition to the polyadenylated nuclear RNA (PAN), a long non-coding RNA encoded by the oncogenic DNA virus Kaposi's sarcoma-associated herpesvirus (KSHV), triggering viral lytic reactivation from latency. Mutagenesis of ac⁴C sites in PAN RNA in the context of KSHV infection abolishes PAN ac⁴C modifications, downregulates the expression of viral lytic genes and reduces virion production. NAT10 knockdown or mutagenesis eras2023-01-01T00:00:00Z2023 Oct2026-05-29T00:39:46.336Z2025-02-18T23:52:45.855ZS-EPMC105648943781677110.1038/s41467-023-42135-3