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The mechanisms of flaviviruses evade host translational shutoff are largely unknown. Here, it is found that Japanese encephalitis virus (JEV) can adopt cap\u2010independent (CI) translation to escape the host translational shutoff. Furthermore, the elements DB2 and sHP\u2010SL within 3\u2032UTR are involved in the regulation of CI translation, which is conserved in the genus\n                    <jats:italic>Orthoflavivirus<\/jats:italic>\n                    . By RNA affinity purification and mass spectrometry analysis, cellular DEAD\u2010box protein 3 (DDX3) and poly(A)\u2010binding protein 1 (PABP1) are identified as key factors in regulating CI translation of JEV via their interactions with DB2 and sHP\u2010SL RNA structures. Mechanistically, it is revealed that DDX3 binds to both 5\u2032UTR and 3\u2032UTR of the JEV genome to establish a closed\u2010loop architecture and recruit eIF4G\/eIF4A to form the DDX3\/PABP1\/eIF4G\/eIF4A tetrameric complex via its interaction with PABP1, thereby recruiting the ribosomal 43S preinitiation complex (PIC) to the 5\u2032\u2010end of the JEV genome to start translation. These findings demonstrate a noncanonical translation strategy employed by JEV and further reveal the regulatory roles of DDX3 and PABP1 in this mechanism. These results expand the knowledge of the translation initiation regulation in flaviviruses under the state of host translational shutoff, which provides a conserved antiviral target against\n                    <jats:italic>orthoflavivirus<\/jats:italic>\n                    .\n                  <\/jats:p>","DOI":"10.1002\/advs.202502493","type":"journal-article","created":{"date-parts":[[2025,5,9]],"date-time":"2025-05-09T02:42:44Z","timestamp":1746758564000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["DDX3 Regulates the Cap\u2010Independent Translation of the Japanese Encephalitis Virus via Its Interactions with PABP1 and the Untranslated Regions of the Viral Genome"],"prefix":"10.1002","volume":"12","author":[{"given":"Chenxi","family":"Li","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine Yangzhou University  Yangzhou Jiangsu 225009 China"},{"name":"Jiangsu Co\u2010innovation Center 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