Ha-VP39 binding to actin and the influence of F-actin on assembly of progeny virions
Ha-VP39 binding to actin and the influence of F-actin on assembly of progeny virions
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DOI：
10.1007/s00705-004-0361-4
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发表时间：
2004-11-01
影响因子：
2.7
通讯作者：
Qi, Y
中科院分区：
文献类型：
--
作者：
Lu, S;Ge, G;Qi, Y
我们提出的证据表明，肌动蛋白是成功组装HaNPV病毒粒子所必需的。Western blot和等温滴定量热法证实，纯化的棉蚜核多角体病毒（HaNPV）核衣壳蛋白Ha-VP39在体外无需辅助即可与肌动蛋白结合。等温滴定量热计测得的δ tah和结合常数K强烈表明，Ha-VP39首先与肌动蛋白结合形成肌动蛋白六聚体复合物，六聚体相互连接形成丝状结构，丝状结构最终缠绕成绳状结构。在含有0.5杯/ml细胞松弛素D （cytochalasin D， CD）的培养基中培养的Hz-AM1细胞中，HaNPV的增殖被完全抑制，以防止肌动蛋白的聚合，而在含有0.1杯/ml CD的培养基中，其产量降低到10（-4）。尽管CD处理的细胞中组装的子代病毒粒子在形态上与正常细胞不同，但CD对肌动蛋白浓度和病毒DNA合成没有显著影响，并且在斑块实验中导致较少的斑块。
We present evidence that actin is necessary for the successful assembly of HaNPV virions. Purified nucleocapsid protein Ha-VP39 of Heliothis armigera nuclear polyhedrosis virus (HaNPV) was found to be able to bind to actin in vitro without assistance, as demonstrated by Western blot and isothermal titration calorimeter. DeltaH and binding constants (K) detected by isothermal titration calorimeter strongly suggested that Ha-VP39 first binds actin to seed the formation of hexamer complex of actin, and the hexamers then link to each other to form filaments, and the filaments finally twist into cable structures. The proliferation of HaNPV was completely inhibited in Hz-AM1 cells cultivated in the medium containing 0.5 mug/ml cytochalasin D (CD) to prevent polymerization of actin, while its yield was reduced to 10(-4) in the presence of 0.1 mug/ml CD. Actin concentration and the viral DNA synthesis were not significantly affected by CD even though the progeny virions assembled in the CD treated cells were morphologically different from normal ones and resulted in fewer plaques in plaque assay.