本文旨在探讨α-actinin 参与丙型肝炎病毒( HCV) 复制的机制。将α-actinin 转染Huh7.5 细胞, 用JFH1 感染, 发现过表达α-actinin 可显著增加HCV RNA 水平及非结构蛋白表达, 感染性HCV 颗粒也同时增多。膜漂浮实验显示,α-actinin 可与HCV 非结构蛋白NS5A 共定位于脂筏。抑制内源性α-actinin 表达,可使复制子细胞内NS5A 表达减少, 且对非离子去污剂敏感而从脂筏脱落。免疫荧光实验显示, NS5A 与内质网标志分子calnexin 核周共定位消失。以上结果提示,α-actinin 可通过影响非结构蛋白与脂筏的关联而参与HCV RNA 的合成, 为临床治疗及研制新型抗HCV 药物提供理论依据和实验基础。

Cellular protein α-actinin has been shown to involve in HCV replication in Huh7 HCV replicon cells . However , it is still elusive howα-actininregulates HCV RNAsynthesis . In this study , we attempted to study the mechanismhowα-actinin facilitates HCVreplication. α- Actinin was transfectedinto JFH1-infected Huh 7.5 cells . The results showed that overexpression of α-actinin significantly increased HCV nonstructural protein expression as well as intracellular and supernatant HCV RNAlevels . More inf ective virus particles were also secreted into media. Membrane flotation analysis indicated that the vast maj ority of both fulllength actinin and NS5A were colocalized in the detergent-resistant membrane ( DRM) fractions . Knockdown of α-actinin altered the membrane association pattern of NS5A. Immunofluorescence microscopy analysis showedthat NS5A was unable to display atypical pattern of cytoplasmic distribution and colocalize with calnexin on endoplasmic reticulum. Taken together , these results indicate that α-actinin regulates HCVreplicationthrough facilitation of NS5A’s associati on with endoplasmic reticulummembrane , and may provide newinsights into fundamental cellular processes and identify novel targets f or antiviral intervention.