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However, the host factors involved in NNV entry remain poorly understood. We identify myosin light chain 3 from marine medaka (\n                    <jats:italic>Oryzias melastigma<\/jats:italic>\n                    ) (MmMYL3) as a novel receptor for red-spotted grouper NNV (RGNNV), facilitating internalization via macropinocytosis. MmMYL3 directly binds the RGNNV capsid protein (CP), which depends on the arm and S domains of CP and the EF-hand2 domain of MmMYL3. In vitro experiments showed that MmMYL3 siRNA, protein, anti-MYL3 antibodies, or the arm domain synthetic peptides blocked RGNNV entry. Moreover, in vivo administration of MmMYL3 protein also inhibited RGNNV infection. Ectopic MmMYL3 expression enabled RGNNV internalization into resistant cells. Notably, MmMYL3 facilitated RGNNV internalization through the macropinocytosis pathway via the IGF1R-Rac1\/Cdc42 axis. Collectively, our findings underscore MYL3\u2019s crucial role in NNV entry and its potential as an antiviral target.\n                  <\/jats:p>","DOI":"10.7554\/elife.104772","type":"journal-article","created":{"date-parts":[[2024,12,17]],"date-time":"2024-12-17T07:25:25Z","timestamp":1734420325000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.104772","source":"Crossref","is-referenced-by-count":5,"title":["Myosin light chain 3 serves as a receptor for nervous necrosis virus entry into host cells via the macropinocytosis pathway"],"prefix":"10.7554","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-6725-4195","authenticated-orcid":true,"given":"Lan","family":"Yao","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0064kty71","id-type":"ROR","asserted-by":"publisher"}],"name":"School of Marine Sciences, Sun Yat-sen University","place":["Zhuhai, 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