ZNF33B Promotes Japanese Encephalitis Virus Infection by Regulating the Stability of M6A-Modified Trim25 to Control the Autophagy Process

Japanese encephalitis virus (JEV) is a neurotropic Flavivirus that causes a substantial threat to human health and livestock; however, the epitranscriptomic mechanisms that support its replication remain poorly defined. Here, we identify a proviral host factor C₂H₂ zinc-finger protein ZNF33B that promotes JEV Infection through coupling N⁶-methyladenosine (m⁶A) RNA modification to Autophagy regulation. Mechanistically, ZNF33B recruits METTL14 to stabilize the METTL3-METTL14 methyltransferase complex, thereby increasing global m⁶A deposition. Multi-omics analyses reveal that ZNF33B selectively binds m⁶A-modified sites within the Antiviral transcript Trim25 (c.1567 and c.1669 bp) to accelerate its decay. We further demonstrate that TRIM25 functions as an E3 ubiquitin Ligase that catalyzes K48-linked ubiquitination of Atg7 at lysines 389 and 423, leading to its proteasomal degradation and ultimately suppressing autophagic flux. In contrast, ZNF33B-mediated Trim25 degradation counteracts its inhibitory effect on Autophagy, creating a favorable environment for viral replication. In vivo, adeno-associated virus (AAV)-mediated ZNF33B delivery increases mouse brain m⁶A levels, decreases TRIM25 expression, elevates Atg7 abundance, exacerbates JEV-induced neuropathology, and accelerates mouse mortality. Together, these findings reveal a previously uncharacterized ZNF33B-m⁶A-TRIM25-autophagy axis that JEV hijacks to evade host Antiviral responses, providing new insights into flaviviral pathogenesis and potential therapeutic targets.