Viral encephalopathy and retinopathy, also known as viral nervous necrosis, is a highly contagious and pathogenic disease that infects more than 120 marine and freshwater fish species, leading to mass mortalities and severe economic losses in the aquaculture industry. The etiological agent responsible, nervous necrosis virus (NNV), belongs to the Nodaviridae family, genus Betanodavirus and manifests as a nonenveloped single-stranded RNA virus (Costa and Thompson, 2016; Munday et al., 2002). The NNV genome is composed of two RNA segments: RNA1 encodes the RNA-dependent RNA polymerase (RDRP), while RNA2 encodes the viral capsid protein (CP), which plays a critical role in the initial interaction with host cell surface receptors, thus facilitating viral entry (Bandín and Souto, 2020; Mori et al., 1992).

Viral entry into host cells is a key determinant of host specificity, tissue tropism, and viral pathogenicity, and involves intricate interactions between viral CPs and host cell receptors (Grove and Marsh, 2011). Viruses utilize sophisticated strategies to attach to one or multiple receptors, enabling them to cross the plasma membrane and access the necessary host cell machinery (Maginnis, 2018). Viral receptors function not only as attachment moieties but also as entry factors, coordinators of viral trafficking, and activators of signaling events (Marsh and Helenius, 2006). Therefore, identifying virus receptors and understanding the mechanism of virus-receptor interaction is critical for better preventing viral diseases. Although NNV exhibits a broad host tropism, only heat shock cognate protein 70 (HSC70), heat shock protein 90ab1 (HSP90ab1), and Nectin1 have been identified as its receptor or co-receptor (Chang and Chi, 2015; Zhang et al., 2020; Zhang et al., 2024). Following NNV CP recognition of the cellular surface receptor, the NNV enters the cell by receptor-mediated endocytosis. Viruses usually take advantage of various endocytic pathways to enter host cells, with clathrin-mediated endocytosis (CME), macropinocytosis, and caveolar/raft-dependent endocytosis being the most extensively studied (Mercer et al., 2010). Previous studies revealed that NNV particles utilize the receptor-mediated CME and macropinocytosis internalization pathways to invade host cells (Liu et al., 2005), with the former facilitated by HSP90ab1 and HSC70 (Zhang et al., 2020). However, the receptor responsible for mediating NNV invasion of host cells through macropinocytosis remains unknown.

Among the diverse cellular factors that can serve as viral receptors, members of the myosin family have emerged as intriguing candidates (He et al., 2022). The myosin family comprises a wide range of actin-associated motor proteins that play a role in a variety of cellular activities, including signal transduction, intracellular transport, cell division, etc. Up to date, more than 40 known classes of myosin have been identified (Foth et al., 2006). Recent studies underscore their significance in various viral infections. For instance, myosin heavy chain 9 has been identified as a crucial factor for various virus infections (Li et al., 2018). Non-muscle myosin heavy chain IIA serves as a functional entry receptor for herpes simplex virus-1 (Arii et al., 2010). Additionally, myosin II light chain activation is essential for influenza A virus cell entry through macropinocytosis (Banerjee et al., 2014).

Myosin light chain 3 (MYL3), a constituent of the myosin light chain family predominantly found in muscle tissues, particularly the heart and skeletal muscles, plays a pivotal role in muscle contraction by forming the myosin complex with myosin heavy chain and facilitating actin filament sliding (Ingles et al., 2005; Zhang et al., 2016). Mutations in the MYL3 gene have been associated with various muscle-related disorders, including cardiomyopathies and skeletal muscle myopathies (Mavilakandy and Ahamed, 2022). However, the role of MYL3 in virus infection remains undiscovered. Huang et al. reported that Epinephelus coioides MYL3 might interact with native NNV CP by proteomic analysis of immunoprecipitation (IP) assay (Huang et al., 2020). In our previous study, we also found that marine medaka (Oryzias melastigma) MYL3 (MmMYL3) is a potential interacting protein of the purified CP (Zhang et al., 2020), yet its involvement in NNV infection, particularly its role in viral invasion, remains uncertain.

In this study, we unveil MmMYL3 as a novel NNV entry receptor, elucidating its facilitation of RGNNV entry into host cells through macropinocytosis, mediated by the IGF1R-Rac1/Cdc42 pathway. These findings offer novel insights into the intricate mechanisms underlying NNV entry and highlight potential targets for the development of antiviral drugs.

NNV, an important fish virus, enters host cells by utilizing a range of host factors that act as receptors and facilitate the process of endocytosis via an orchestrated mechanism. Three host factors, HSP90ab1, HSC70, and Nectin1, have been identified as entry receptors of NNV (Chang and Chi, 2015; Zhang et al., 2020, n.d.); however, disrupting their interaction with cells does only partially hinder viral infection, suggesting that NNV likely utilizes additional, yet unidentified host receptors or co-receptors. To establish infection, NNV utilized receptor-mediated endocytosis to induce internalization of the virus/receptor complex, primarily including CME and macropinocytosis (Liu et al., 2005). Although previous studies demonstrate that HSP90ab1 and HSC70 are responsible for NNV internalization via CME, the receptor and the molecular mechanisms leading to NNV entry into cells via macropinocytosis remain to be determined. In this study, we found that NNV gains entry into host cells via the interaction between CP and its novel receptor MmMYL3. Subsequently, MmMYL3 engages with IGF1R, leading to the activation of the Rac1/Cdc42 pathway, which induces actin rearrangement and facilitates NNV macropinocytosis (Figure 7N).

Numerous data indicate the significance of myosin light chains in the functionality of both cardiac and skeletal muscles. However, little is known about the function of myosin light chains during virus infection (Kamm and Stull, 2011; Sheikh et al., 2016; Yu et al., 2016). Here, we provide several lines of evidence showing that MmMYL3 acts as a novel receptor of NNV. First, ectopic expression of MmMYL3 facilitates internalization of RGNNV, which is mediated by the interaction between the EF-hand2 domain of MmMYL3 and the arm and S domain of CP. Considering that our previous study had reported that MmHSP90ab1, a known NNV receptor, interacted with the linker region of CP (Zhang et al., 2020). Thus, we speculated that NNV might have the ability to utilize multiple receptors simultaneously for cell infection through these receptors binding to the different domains of CP. Furthermore, interference with the MmMYL3 and CP interaction with abs against MYL3, MmMYL3 fusion protein, and CP arm domain peptides significantly blocks NNV entry, providing a potential antiviral strategy against NNV infection. In addition to these in vitro findings, our in vivo experiments further confirm the physiological relevance of MmMYL3 in viral pathogenesis and highlight its potential as a therapeutic target in aquaculture. More importantly, virus-resistant HEK293T cells are rendered susceptible to RGNNV infection by overexpression of MmMYL3 alone. The development of MmMYL3-overexpressing HEK293T cell model offers a valuable platform for investigating viral infection mechanisms and conducting drug screening studies. All these results suggested that MmMYL3 is a novel receptor that mediates NNV entry. We also found that the interaction of MYL3 with CP was conserved across NNV-sensitive fish. Interestingly, NNV exhibits a broad species tropism, capable of infecting diverse cell lines and various animal species examined thus far (Munday et al., 2002). This phenomenon may be attributed in part to the robust conservation of MYL3. To the best of our knowledge, this is the first study to provide evidence that MYL3 functions as a functional entry receptor for virus.

Nodaviridae viruses are divided into two genera: Alphanodavirus and Betanodavirus. Alphanodaviruses typically infect insects, whereas Betanodaviruses primarily infect fish (Yong et al., 2017). In recent years, a novel Alphanodavirus, provisionally named CMNV, has emerged, causing viral covert mortality disease in shrimp and also affecting farmed marine fish (Wang et al., 2021; Zhang et al., 2014). Given the susceptibility of marine fish to CMNV, we investigated the possibility that both Alphanodavirus and Betanodavirus utilize MYL3 as a common receptor for cellular entry. Sequence comparison results revealed a low homology between the CP sequences of CMNV and RGNNV. Additionally, our data indicated that the CP of CMNV did not interact with MmMYL3. These findings suggest that MmMYL3 may function as a receptor specific to Betanodavirus, rather than being a common receptor for both viral genera. Receptor recognition is typically highly specific, and the binding interactions between viral proteins and host receptors often depend on the structural compatibility between the viral capsid/viral envelope and the host receptor. Since MYL3 does not interact with CMNV, a virus more closely related to NNV, it is less likely to function as a receptor for viruses that are more distantly related to NNV.

Previous reports have highlighted the dependence of the NNV entry process on both CME and macropinocytosis pathways (Liu et al., 2005). We had demonstrated that NNV can use MmHSP90ab1 and MmHSC70 to enter cells via the CME pathway (Chang and Chi, 2015; Zhang et al., 2020). In contrast to these receptors, our findings revealed that MmMYL3 promotes NNV internalization via macropinocytosis. Macropinocytosis, characterized as a transient, growth factor-induced, actin-dependent endocytic pathway, facilitates the internalization of fluid and membrane into large vacuoles (Mercer and Helenius, 2009). Mounting evidence underscores its pivotal role as an endocytic route exploited by numerous viruses for invading host cells (Huang et al., 2008; Locker et al., 2000; Mercer and Helenius, 2008). Upon binding to the cell membrane, viruses trigger the activation of RTKs or other signaling molecules such as Rac1, Cdc42, and Pak1, initiating intricate intracellular signaling cascades that lead to alterations in actin filament dynamics and induction of plasma membrane ruffling (Bar-Sagi and Feramisco, 1986; Bar-Sagi et al., 1987; Letizia Lanzetti APLAS and PPDF, 2004). RTKs, well-known upstream regulatory factors of macropinocytosis, are involved in the entry process of various viruses (Amyere et al., 2000; Nicola et al., 2005). To evaluate the significance of RTKs in NNV entry, we conducted a screening of small molecules targeting the activation of specific RTKs. Our findings revealed that AZD-3463 and Picropodophyllin, two inhibitors of IGF1R, could impede MmMYL3-mediated RGNNV entry. IGF1R is a transmembrane RTK involved in mediating cellular responses to IGFs and plays a significant role in cellular processes such as growth, proliferation, and survival (Hakuno and Takahashi, 2018; Pellegrino et al., 2024). Recent studies have revealed an intriguing connection between IGF1R signaling and viral entry mechanisms, particularly through macropinocytosis (Józefiak et al., 2021; Stewart et al., 2021). Our findings revealed direct binding between MmMYL3 and MmIGF1R, indicating that MmMYL3 might participate in the macropinocytosis-mediated NNV entry through its interaction with MmIGF1R. Previous study had identified IGF1R as an entry receptor for respiratory syncytial virus (Griffiths et al., 2020). Consider the conservation of IGF1R, we investigated whether it could serve as a potential receptor for NNV. However, our analysis indicated that MmIGF1R did not directly bind to CP, indicating that MmIGF1R might not be the receptor for NNV.

To further explore how MmIGF1R promotes macropinocytosis and NNV internalization, we investigate the effect of MmIGF1R downstream signaling on NNV infection. IGF1R can activate GTPases, such as Cdc42 and Rac1, leading to rapid actin reorganization and membrane ruffling, key processes in initiating macropinocytosis (Dam et al., 2017; Manara et al., 2016). Increasing evidence has demonstrated that Cdc42 and Rac1 are essential for the entry of various viruses (Lu et al., 2023; Swaine and Dittmar, 2015; Zhang et al., 2021). We found that both MmRac1 and MmCdc42 interact with the intracellular domain of MmIGF1R, and inhibition of MmRac1 and MmCdc42 significantly impaired RGNNV entry. Conversely, overexpression of MmRac1 or MmCdc42 enhanced NNV internalization. Actin filaments play a central role in driving the membrane dynamics necessary for the formation and internalization of macropinosomes during macropinocytosis. We found that there was a notable reorganization of F-actin filaments, accompanied by the presence of actin-driven membrane protrusions that co-localized with virions on the surface of RGNNV-infected cells. All these results indicated that cytosis MmIGF1R regulates Rac1/Cdc42 signaling to promote macropinocytosis during NNV infection. While previous studies have suggested that NNV enters host cells via macropinocytosis, our study was the first to provide a detailed mechanistic understanding of this process. The identification of MYL3 as a novel receptor for RGNNV entry, coupled with its role in activating the IGF1R-Rac1/Cdc42 axis to drive macropinocytosis, opens new avenues for therapeutic intervention in aquaculture. MYL3, as a crucial receptor for viral entry, could be inhibited through the development of small molecules, peptides, or monoclonal antibodies that block its interaction with the virus. Additionally, IGF1R inhibitors could prevent macropinocytosis and viral entry by disrupting downstream signaling pathways. Screening libraries of small molecules and natural products for potential inhibitors and testing them in aquaculture models would help evaluate their efficacy. Combining MYL3 and IGF1R inhibitors might offer a synergistic approach to prevent viral infection. These strategies could significantly reduce viral loads and improve fish survival rates, addressing the ongoing viral challenges in aquaculture.

Overall, MmMYL3 was identified as a novel functional receptor for RGNNV. Moreover, MmMYL3 was involved in and facilitated RGNNV internalization through the macropinocytosis pathway via the IGF1R-Rac1/Cdc42 axis. Our findings shed new light on the molecular mechanism of RGNNV entry and provide a new perspective for the development of antiviral therapies.

While our study provides strong evidence that MmMYL3 serves as a receptor for RGNNV in vitro, one limitation of our research is the lack of in vivo experiments using MmMYL3 knockout marine medaka. Conducting these knockout studies would further validate the role of MmMYL3 in NNV infection in a whole organism context. Unlike well-established model organisms like zebrafish or mice, gene editing in marine medaka is fraught with difficulties, including technical complexity, precise embryo microinjection, and the potential for off-target effects with CRISPR/Cas9 technology. The process is time-consuming, requiring multiple breeding generations and thorough molecular screening to establish homozygous knockout lines. Future research aimed at developing these knockout models will greatly benefit from advances in gene editing technologies and standardized protocols tailored to marine medaka.

All data generated or analysed during this study are included in the manuscript, figures, figure supplements and source data files. source data files have been provided for Figures 1–7.

1. 1. Amyere M
   2. Payrastre B
   3. Krause U
   4. Van Der Smissen P
   5. Veithen A
   6. Courtoy PJ

   (2000) Constitutive macropinocytosis in oncogene-transformed fibroblasts depends on sequential permanent activation of phosphoinositide 3-kinase and phospholipase C

   Molecular Biology of the Cell 11:3453–3467.

   https://doi.org/10.1091/mbc.11.10.3453

   • PubMed
   • Google Scholar
2. 1. Arii J
   2. Goto H
   3. Suenaga T
   4. Oyama M
   5. Kozuka-Hata H
   6. Imai T
   7. Minowa A
   8. Akashi H
   9. Arase H
   10. Kawaoka Y
   11. Kawaguchi Y

   (2010) Non-muscle myosin IIA is a functional entry receptor for herpes simplex virus-1

   Nature 467:859–862.

   https://doi.org/10.1038/nature09420

   • PubMed
   • Google Scholar
3. 1. Bandín I
   2. Souto S

   (2020) Betanodavirus and VER disease: A 30-year research review

   Pathogens 9:1–46.

   https://doi.org/10.3390/pathogens9020106

   • PubMed
   • Google Scholar
4. 1. Banerjee I
   2. Miyake Y
   3. Nobs SP
   4. Schneider C
   5. Horvath P
   6. Kopf M
   7. Matthias P
   8. Helenius A
   9. Yamauchi Y

   (2014) Influenza A virus uses the aggresome processing machinery for host cell entry

   Science 346:473–477.

   https://doi.org/10.1126/science.1257037

   • PubMed
   • Google Scholar
5. 1. Bar-Sagi D
   2. Feramisco JR

   (1986) Induction of membrane ruffling and fluid-phase pinocytosis in quiescent fibroblasts by ras proteins

   Science 233:1061–1068.

   https://doi.org/10.1126/science.3090687

   • PubMed
   • Google Scholar
6. 1. Bar-Sagi D
   2. McCormick F
   3. Milley RJ
   4. Feramisco JR

   (1987) Inhibition of cell surface ruffling and fluid-phase pinocytosis by microinjection of anti-ras antibodies into living cells

   Journal of Cellular Physiology 133:69–73.

   https://doi.org/10.1002/jcp.1041330414

   • Google Scholar
7. 1. Chang JS
   2. Chi SC

   (2015) GHSC70 is involved in the cellular entry of nervous necrosis virus

   Journal of Virology 89:61–70.

   https://doi.org/10.1128/JVI.02523-14

   • PubMed
   • Google Scholar
8. 1. Costa JZ
   2. Thompson KD

   (2016) Understanding the interaction between Betanodavirus and its host for the development of prophylactic measures for viral encephalopathy and retinopathy

   Fish & Shellfish Immunology 53:35–49.

   https://doi.org/10.1016/j.fsi.2016.03.033

   • PubMed
   • Google Scholar
9. 1. Dam DHM
   2. Wang XQ
   3. Sheu S
   4. Vijay M
   5. Shipp D
   6. Miller L
   7. Paller AS

   (2017) Ganglioside GM3 mediates glucose-induced suppression of IGF-1 Receptor-Rac1 activation to inhibit keratinocyte motility

   The Journal of Investigative Dermatology 137:440–448.

   https://doi.org/10.1016/j.jid.2016.09.028

   • PubMed
   • Google Scholar
10. 1. Delpeut S
    2. Sisson G
    3. Black KM
    4. Richardson CD

    (2017) Measles virus enters breast and colon cancer cell lines through a PVRL4-mediated macropinocytosis pathway

    Journal of Virology 91:e02191-16.

    https://doi.org/10.1128/JVI.02191-16

    • PubMed
    • Google Scholar
11. 1. Dong S
    2. Kang M
    3. Wu X
    4. Ye T

    (2014) Development of a promising fish model (Oryzias melastigma) for assessing multiple responses to stresses in the marine environment

    BioMed Research International 2014:563131.

    https://doi.org/10.1155/2014/563131

    • PubMed
    • Google Scholar
12. 1. Foth BJ
    2. Goedecke MC
    3. Soldati D

    (2006) New insights into myosin evolution and classification

    PNAS 103:3681–3686.

    https://doi.org/10.1073/pnas.0506307103

    • PubMed
    • Google Scholar
13. 1. Griffiths CD
    2. Bilawchuk LM
    3. McDonough JE
    4. Jamieson KC
    5. Elawar F
    6. Cen Y
    7. Duan W
    8. Lin C
    9. Song H
    10. Casanova JL
    11. Ogg S
    12. Jensen LD
    13. Thienpont B
    14. Kumar A
    15. Hobman TC
    16. Proud D
    17. Moraes TJ
    18. Marchant DJ

    (2020) IGF1R is an entry receptor for respiratory syncytial virus

    Nature 583:615–619.

    https://doi.org/10.1038/s41586-020-2369-7

    • PubMed
    • Google Scholar
14. 1. Grove J
    2. Marsh M

    (2011) The cell biology of receptor-mediated virus entry

    The Journal of Cell Biology 195:1071–1082.

    https://doi.org/10.1083/jcb.201108131

    • PubMed
    • Google Scholar
15. 1. Hakuno F
    2. Takahashi SI

    (2018) 40 years OF IGF1: IGF1 receptor signaling pathways

    Journal of Molecular Endocrinology 61:T69–T86.

    https://doi.org/10.1530/JME-17-0311

    • Google Scholar
16. 1. He J
    2. Qiu Y
    3. Tan L
    4. Duan D
    5. Yuan X
    6. Yang L
    7. Wang A

    (2022) Understanding the key functions of Myosins in viral infection

    Biochemical Society Transactions 50:597–607.

    https://doi.org/10.1042/BST20211239

    • PubMed
    • Google Scholar
17. 1. Huang CY
    2. Lu TY
    3. Bair CH
    4. Chang YS
    5. Jwo JK
    6. Chang W

    (2008) A novel cellular protein, VPEF, facilitates vaccinia virus penetration into HeLa cells through fluid phase endocytosis

    Journal of Virology 82:7988–7999.

    https://doi.org/10.1128/JVI.00894-08

    • PubMed
    • Google Scholar
18. 1. Huang PY
    2. Hsiao HC
    3. Wang SW
    4. Lo SF
    5. Lu MW
    6. Chen LL

    (2020) Screening for the proteins that can interact with grouper nervous necrosis virus capsid protein

    Viruses 12:1–20.

    https://doi.org/10.3390/v12090985

    • PubMed
    • Google Scholar
19. 1. Ingles J
    2. Doolan A
    3. Chiu C
    4. Seidman J
    5. Seidman C
    6. Semsarian C

    (2005) Compound and double mutations in patients with hypertrophic cardiomyopathy: implications for genetic testing and counselling

    Journal of Medical Genetics 42:2–7.

    https://doi.org/10.1136/jmg.2005.033886

    • PubMed
    • Google Scholar
20. 1. Jia KT
    2. Wu YY
    3. Liu ZY
    4. Mi S
    5. Zheng YW
    6. He J
    7. Weng SP
    8. Li SC
    9. He JG
    10. Guo CJ

    (2013) Mandarin fish caveolin 1 interaction with major capsid protein of infectious spleen and kidney necrosis virus and its role in early stages of infection

    Journal of Virology 87:3027–3038.

    https://doi.org/10.1128/JVI.00552-12

    • PubMed
    • Google Scholar
21. 1. Jia P
    2. Jia KT
    3. Yi MS

    (2015) Complete genome sequence of a fish nervous necrosis virus isolated from sea perch (Lateolabrax japonicus) in China

    Genome Announcements 3:455643.

    https://doi.org/10.1128/genomeA.00048-15

    • PubMed
    • Google Scholar
22. 1. Józefiak A
    2. Larska M
    3. Pomorska-Mól M
    4. Ruszkowski JJ

    (2021) The IGF-1 signaling pathway in viral infections

    Viruses 13:1–11.

    https://doi.org/10.3390/v13081488

    • PubMed
    • Google Scholar
23. 1. Kamm KE
    2. Stull JT

    (2011) Signaling to myosin regulatory light chain in sarcomeres

    The Journal of Biological Chemistry 286:9941–9947.

    https://doi.org/10.1074/jbc.R110.198697

    • PubMed
    • Google Scholar
24. 1. Le Y
    2. Li Y
    3. Jin Y
    4. Jia P
    5. Jia K
    6. Yi M

    (2017) Establishment and characterization of a brain cell line from sea perch, Lateolabrax japonicus

    In Vitro Cellular & Developmental Biology - Animal 53:834–840.

    https://doi.org/10.1007/s11626-017-0185-7

    • Google Scholar
25. 1. Letizia Lanzetti APLAS
    2. PPDF

    (2004) Rab5 is a signalling GTPases

    Nature 429:309–314.

    https://doi.org/10.1038/nature02542

    • Google Scholar
26. 1. Li L
    2. Xue B
    3. Sun W
    4. Gu G
    5. Hou G
    6. Zhang L
    7. Wu C
    8. Zhao Q
    9. Zhang Y
    10. Zhang G
    11. Hiscox JA
    12. Nan Y
    13. Zhou EM

    (2018) Recombinant MYH9 protein C-terminal domain blocks porcine reproductive and respiratory syndrome virus internalization by direct interaction with viral glycoprotein 5

    Antiviral Research 156:10–20.

    https://doi.org/10.1016/j.antiviral.2018.06.001

    • PubMed
    • Google Scholar
27. 1. Liu W
    2. Hsu C-H
    3. Hong Y-R
    4. Wu S-C
    5. Wang C-H
    6. Wu Y-M
    7. Chao C-B
    8. Lin C-S

    (2005) Early endocytosis pathways in SSN-1 cells infected by dragon grouper nervous necrosis virus

    The Journal of General Virology 86:2553–2561.

    https://doi.org/10.1099/vir.0.81021-0

    • PubMed
    • Google Scholar
28. 1. Locker JK
    2. Kuehn A
    3. Schleich S
    4. Rutter G
    5. Hohenberg H
    6. Wepf R
    7. Griffiths G

    (2000) Entry of the two infectious forms of vaccinia virus at the plasma membane is signaling-dependent for the IMV but not the EEV

    Molecular Biology of the Cell 11:2497–2511.

    https://doi.org/10.1091/mbc.11.7.2497

    • PubMed
    • Google Scholar
29. 1. Lu ZZ
    2. Sun C
    3. Zhang X
    4. Peng Y
    5. Wang Y
    6. Zeng Y
    7. Zhu N
    8. Yuan Y
    9. Zeng MS

    (2023) Neuropilin 1 is an entry receptor for KSHV infection of mesenchymal stem cell through TGFBR1/2-mediated macropinocytosis

    Science Advances 9:eadg1778.

    https://doi.org/10.1126/sciadv.adg1778

    • PubMed
    • Google Scholar
30. 1. Lv W
    2. Jiang W
    3. Luo H
    4. Tong Q
    5. Niu X
    6. Liu X
    7. Miao Y
    8. Wang J
    9. Guo Y
    10. Li J
    11. Zhan X
    12. Hou Y
    13. Peng Y
    14. Wang J
    15. Zhao S
    16. Xu Z
    17. Zuo B

    (2022) Long noncoding RNA lncMREF promotes myogenic differentiation and muscle regeneration by interacting with the Smarca5/p300 complex

    Nucleic Acids Research 50:10733–10755.

    https://doi.org/10.1093/nar/gkac854

    • PubMed
    • Google Scholar
31. 1. Maginnis MS

    (2018) Virus-receptor interactions: the key to cellular invasion

    Journal of Molecular Biology 430:2590–2611.

    https://doi.org/10.1016/j.jmb.2018.06.024

    • PubMed
    • Google Scholar
32. 1. Manara MC
    2. Terracciano M
    3. Mancarella C
    4. Sciandra M
    5. Guerzoni C
    6. Pasello M
    7. Grilli A
    8. Zini N
    9. Picci P
    10. Colombo MP
    11. Morrione A
    12. Scotlandi K

    (2016) CD99 triggering induces methuosis of Ewing sarcoma cells through IGF-1R/RAS/Rac1 signaling

    Oncotarget 7:79925–79942.

    https://doi.org/10.18632/oncotarget.13160

    • PubMed
    • Google Scholar
33. 1. Marsh M
    2. Helenius A

    (2006) Virus entry: open sesame

    Cell 124:729–740.

    https://doi.org/10.1016/j.cell.2006.02.007

    • PubMed
    • Google Scholar
34. 1. Mavilakandy A
    2. Ahamed H

    (2022) Mutation of the MYL3 gene in a patient with mid-ventricular obstructive hypertrophic cardiomyopathy

    BMJ Case Reports 15:1–5.

    https://doi.org/10.1136/bcr-2021-244573

    • PubMed
    • Google Scholar
35. 1. Mercer J
    2. Helenius A

    (2008) Vaccinia virus uses macropinocytosis and apoptotic mimicry to enter host cells

    Science 320:531–535.

    https://doi.org/10.1126/science.1155164

    • PubMed
    • Google Scholar
36. 1. Mercer J
    2. Helenius A

    (2009) Virus entry by macropinocytosis

    Nature Cell Biology 11:510–520.

    https://doi.org/10.1038/ncb0509-510

    • PubMed
    • Google Scholar
37. 1. Mercer J
    2. Schelhaas M
    3. Helenius A

    (2010) Virus entry by endocytosis

    Annual Review of Biochemistry 79:803–833.

    https://doi.org/10.1146/annurev-biochem-060208-104626

    • PubMed
    • Google Scholar
38. 1. Mori KI
    2. Nakai T
    3. Muroga K
    4. Arimoto M
    5. Mushiake K
    6. Furusawa I

    (1992) Properties of a new virus belonging to nodaviridae found in larval striped jack (Pseudocaranx dentex) with nervous necrosis

    Virology 187:368–371.

    https://doi.org/10.1016/0042-6822(92)90329-N

    • Google Scholar
39. 1. Munday BL
    2. Kwang J
    3. Moody N

    (2002) Betanodavirus infections of teleost fish: A review

    Journal of Fish Diseases 25:127–142.

    https://doi.org/10.1046/j.1365-2761.2002.00350.x

    • Google Scholar
40. 1. Nicola AV
    2. Hou J
    3. Major EO
    4. Straus SE

    (2005) Herpes simplex virus type 1 enters human epidermal keratinocytes, but not neurons, via a pH-dependent endocytic pathway

    Journal of Virology 79:7609–7616.

    https://doi.org/10.1128/JVI.79.12.7609-7616.2005

    • PubMed
    • Google Scholar
41. 1. Pellegrino M
    2. Secli V
    3. D’Amico S
    4. Petrilli LL
    5. Caforio M
    6. Folgiero V
    7. Tumino N
    8. Vacca P
    9. Vinci M
    10. Fruci D
    11. de Billy E

    (2024) Manipulating the tumor immune microenvironment to improve cancer immunotherapy: IGF1R, a promising target

    Frontiers in Immunology 15:1356321.

    https://doi.org/10.3389/fimmu.2024.1356321

    • PubMed
    • Google Scholar
42. 1. Sheikh F
    2. Lyon RC
    3. Chen J
    4. Jolla L

    (2016) HHS public access

    Gene 569:14–20.

    https://doi.org/10.1016/j.gene.2015.06.027

    • Google Scholar
43. 1. Stewart CM
    2. Phan A
    3. Bo Y
    4. LeBlond ND
    5. Smith TKT
    6. Laroche G
    7. Giguère PM
    8. Fullerton MD
    9. Pelchat M
    10. Kobasa D
    11. Côté M

    (2021) Ebola virus triggers receptor tyrosine kinase-dependent signaling to promote the delivery of viral particles to entry-conducive intracellular compartments

    PLOS Pathogens 17:e1009275.

    https://doi.org/10.1371/journal.ppat.1009275

    • PubMed
    • Google Scholar
44. 1. Swaine T
    2. Dittmar MT

    (2015) CDC42 use in viral cell entry processes by RNA Viruses

    Viruses 7:6526–6536.

    https://doi.org/10.3390/v7122955

    • PubMed
    • Google Scholar
45. 1. Wang C
    2. Liu S
    3. Tang KFJ
    4. Zhang Q

    (2021) Natural infection of covert mortality nodavirus affects Zebrafish (Danio rerio)

    Journal of Fish Diseases 44:1315–1324.

    https://doi.org/10.1111/jfd.13390

    • PubMed
    • Google Scholar
46. 1. Yong CY
    2. Yeap SK
    3. Omar AR
    4. Tan WS

    (2017) Advances in the study of nodavirus

    PeerJ 5:e3841.

    https://doi.org/10.7717/peerj.3841

    • PubMed
    • Google Scholar
47. 1. Yu H
    2. Chakravorty S
    3. Song W
    4. Ferenczi MA

    (2016) Phosphorylation of the regulatory light chain of myosin in striated muscle: methodological perspectives

    European Biophysics Journal 45:779–805.

    https://doi.org/10.1007/s00249-016-1128-z

    • PubMed
    • Google Scholar
48. 1. Zhang Q
    2. Liu Q
    3. Liu S
    4. Yang H
    5. Liu S
    6. Zhu L
    7. Yang B
    8. Jin J
    9. Ding L
    10. Wang X
    11. Liang Y
    12. Wang Q
    13. Huang J

    (2014) A new nodavirus is associated with covert mortality disease of shrimp

    The Journal of General Virology 95:2700–2709.

    https://doi.org/10.1099/vir.0.070078-0

    • PubMed
    • Google Scholar
49. 1. Zhang C
    2. Wang J
    3. Wang G
    4. Ji Z
    5. Hou L
    6. Liu Z
    7. Chao T

    (2016) Molecular cloning and mRNA expression analysis of sheep MYL3 and MYL4 genes

    Gene 577:209–214.

    https://doi.org/10.1016/j.gene.2015.11.041

    • PubMed
    • Google Scholar
50. 1. Zhang W
    2. Jia K
    3. Jia P
    4. Xiang Y
    5. Lu X
    6. Liu W
    7. Yi M

    (2020) Marine medaka heat shock protein 90ab1 is a receptor for red-spotted grouper nervous necrosis virus and promotes virus internalization through clathrin-mediated endocytosis

    PLOS Pathogens 16:e1008668.

    https://doi.org/10.1371/journal.ppat.1008668

    • PubMed
    • Google Scholar
51. 1. Zhang F
    2. Liu Y
    3. You Q
    4. Yang E
    5. Liu B
    6. Wang H
    7. Xu S
    8. Nawaz W
    9. Chen D
    10. Wu Z

    (2021) NSC23766 and Ehop016 suppress herpes simplex virus-1 replication by inhibiting Rac1 activity

    Biological & Pharmaceutical Bulletin 44:1263–1271.

    https://doi.org/10.1248/bpb.b21-00054

    • PubMed
    • Google Scholar
52. 1. Zhang YY
    2. Liang R
    3. Wang SJ
    4. Ye ZW
    5. Wang TY
    6. Chen M
    7. Liu J
    8. Na L
    9. Yang YL
    10. Yang YB
    11. Yuan S
    12. Yin X
    13. Cai XH
    14. Tang YD

    (2022a) SARS-CoV-2 hijacks macropinocytosis to facilitate its entry and promote viral spike-mediated cell-to-cell fusion

    The Journal of Biological Chemistry 298:102511.

    https://doi.org/10.1016/j.jbc.2022.102511

    • PubMed
    • Google Scholar
53. 1. Zhang W
    2. Weng J
    3. Yao L
    4. Jia P
    5. Yi M
    6. Jia K

    (2022b) Nectin4 antagonises type I interferon production by targeting TRAF3 for autophagic degradation and disrupting TRAF3-TBK1 complex formation

    International Journal of Biological Macromolecules 218:654–664.

    https://doi.org/10.1016/j.ijbiomac.2022.07.151

    • Google Scholar
54. 1. Zhang Z
    2. Xing J
    3. Tang X
    4. Sheng X
    5. Chi H
    6. Zhan W

    (2024) Nectin1 is a pivotal host factor involved in attachment and entry of red-spotted grouper nervous necrosis virus in the early stages of the viral life cycle

    Journal of Virology 98:1–27.

    https://doi.org/10.1128/jvi.00901-24

    • Google Scholar

Author details

1. Lan Yao

   1. School of Marine Sciences, Sun Yat-sen University, Zhuhai, China
   2. Southern Marine Science and Engineering Guangdong Laboratory, Zhuhai, China
   3. Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, Guangzhou, China

   Contribution

   Data curation, Formal analysis, Validation, Investigation, Visualization, Methodology, Writing – original draft, Writing – review and editing

   Contributed equally with

   Wanwan Zhang

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0009-0003-6725-4195

2. Wanwan Zhang

   1. School of Marine Sciences, Sun Yat-sen University, Zhuhai, China
   2. Southern Marine Science and Engineering Guangdong Laboratory, Zhuhai, China
   3. Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, Guangzhou, China

   Contribution

   Formal analysis, Funding acquisition, Validation, Investigation, Visualization, Methodology, Writing – original draft, Writing – review and editing

   Contributed equally with

   Lan Yao

   Competing interests

   No competing interests declared

3. Xiaogang Yang

   1. School of Marine Sciences, Sun Yat-sen University, Zhuhai, China
   2. Southern Marine Science and Engineering Guangdong Laboratory, Zhuhai, China
   3. Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, Guangzhou, China

   Contribution

   Formal analysis, Investigation, Methodology

   Competing interests

   No competing interests declared

4. Meisheng Yi

   1. School of Marine Sciences, Sun Yat-sen University, Zhuhai, China
   2. Southern Marine Science and Engineering Guangdong Laboratory, Zhuhai, China
   3. Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, Guangzhou, China

   Contribution

   Conceptualization, Supervision, Funding acquisition, Project administration, Writing – review and editing

   For correspondence

   yimsh@mail.sysu.edu.cn

   Competing interests

   No competing interests declared

5. Kuntong Jia

   1. School of Marine Sciences, Sun Yat-sen University, Zhuhai, China
   2. Southern Marine Science and Engineering Guangdong Laboratory, Zhuhai, China
   3. Guangdong Provincial Key Laboratory of Marine Resources and Coastal Engineering, Guangzhou, China

   Contribution

   Conceptualization, Supervision, Funding acquisition, Investigation, Writing – original draft, Project administration, Writing – review and editing

   For correspondence

   jiakt3@mail.sysu.edu.cn

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0001-7674-9454

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