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oai:figshare.com:article/34017568 2026-09-28T17:52:08Z category_12 category_24 category_272 category_39 category_734 category_69 category_106 category_132 category_134 portal_5 item_type_1 month_year_09_2026

Palmitoylation of the SVCV G protein is essential for viral budding. Chen Li (54018) Yan Zhang (8098) Yujun Zhang (118792) Yan Gao (93649) Yuanan Lu (324035) Shengbo Cao (105106) Jing Ye (94291) Xueqin Liu (494581) Cell Biology Evolutionary Biology Environmental Sciences not elsewhere classified Ecology Biological Sciences not elsewhere classified Inorganic Chemistry Science Policy Infectious Diseases Virology significant economic impacts pathogenicity remain unclear modification enhances stability hold broader significance employed spring viremia competitive peptide targeting cell membrane localization 20 &# 176 molecular mechanisms underlying affects rhabdovirus infections temperature adaptive evolution rhabdovirus prevalent mechanism underlying xlink "> svcv glycoprotein svcv ), similar mechanism palmitic acid numerous rhabdoviruses high infectivity global aquaculture g protein g ). findings elucidate dependent infectivity crucial factor carp virus c ), aquatic rhabdoviruses acyltransferase zdhhc15a (A-B) ZF4 cells were transfected with the plasmid encoding SVCV G for 24h at 28 °C and 16 °C. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 1 h, 2 h, and 4 h). The protein levels of SVCV-G in the cell lysate were measured by Western blotting. (C) ZF4 cells were transfected with the plasmid encoding SVCV G(C484/485A) for 24h at 28 °C and 16 °C. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 1 h, 2 h, and 4 h). The protein level of SVCV-G in the cell lysate was measured by Western blotting. (D) ZF4 cells were transfected with the plasmid encoding SVCV G and were then treated with 2-BP (25µM, 50 µM, and 100 µM) or DMSO for 24 h at 16 °C. The SVCV G protein levels were measured by Western blotting. (E) ZF4 cells were transfected with EGFP-SVCV G plasmid for 24 h at 16°C and 28°C. Cell membranes were stained with Dil and nuclei with DAPI. Scale bars,10 μm. (F) ZF4 cells expressing EGFP-SVCV G were treated with control solvent or 2-BP (50 μM) for 24 h at 16°C. Cell membranes were stained with Dil and nuclei with DAPI. Scale bars,10 μm. (G) ZF4 cells were transfected with EGFP-SVCV G or the mutants for 24 h at 16°C. Cell membranes were stained with Dil and nuclei with DAPI. Scale bars,10 μm. (H) ZF4 cells were co-transfected with SVCV G and SVCV M for 24 h at 16°C and 28 °C. The protein levels of SVCV-M and SVCV-G in the cell lysate and culture supernatants were measured by Western blotting. (I) ZF4 cells were transfected with plasmids expressing SVCV G/the mutants and SVCV M at 16 °C and subsequently mock treated (SVCV G/the mutants and SVCV M groups) or treated with 50 mM 2-BP (SVCV G 2-BP group). The protein levels of SVCV-M and SVCV-G in the cell lysate and culture supernatants were measured by Western blotting. (J) ZF4 cells were infected with SVCV (MOI = 10) under different temperature conditions. Following 12 hours post-infection, the cells were fixed with glutaraldehyde and processed for transmission electron microscopy to observe SVCV budding. Representative images are shown in (J), with scale bars of 5 μm and 1 μm. (K) The budding efficiency of SVCV in ZF4 cells cultured at different temperatures was assessed by calculating the ratio of intracellular and extracellular infectious virus titers to the total infectious titer at 12 hours post-infection. Data was presented as mean ± SD from three independent experiments. Statistical analysis was performed using Student’s t-test between different groups and the asterisk (*) indicates significant differences between groups*p < 0.05, **p < 0.01, ***p < 0.001. 2026-09-28T17:51:51Z Image Figure 10.1371/journal.ppat.1014654.g002 https://figshare.com/articles/figure/_p_Palmitoylation_of_the_SVCV_G_protein_is_essential_for_viral_budding_p_/34017568 CC BY 4.0