VACCINIA VIRUS-REPLICATION .1. REQUIREMENT FOR THE HOST-CELL NUCLEUS
VACCINIA VIRUS-REPLICATION .1. REQUIREMENT FOR THE HOST-CELL NUCLEUS
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DOI：
10.1128/jvi.29.2.705-715.1979
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发表时间：
1979-01-01
影响因子：
5.4
通讯作者：
KATES, JR
中科院分区：
文献类型：
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作者：
HRUBY, DE;GUARINO, LA;KATES, JR
细胞松弛素B诱导的去核技术被用来检查牛痘病毒在几种哺乳动物细胞系中在宿主细胞核不存在的情况下复制的能力。从BSC-40 [非洲绿色猴肾]、CVC和L [小鼠成纤维细胞]细胞制备的病毒感染的去核细胞（胞质体）不能产生感染性子代病毒。在BSC-40细胞中详细研究了这种明显的核参与的性质。旨在最大限度地提高细胞质完整性和寿命的调制，如降低生长温度和初始感染复数，并没有改善病毒在细胞质中的生长。十二烷基硫酸钠-聚丙烯酰胺凝胶分析的[35 S]蛋氨酸脉冲标记的蛋白质在牛痘病毒感染的细胞质中合成表明，早期和晚期的病毒基因产物表达在高水平和适当的时间序列。牛痘病毒胞质DNA合成，测定[3 H]胸苷掺入，在感染后3小时达到峰值，是70-90%的控制水平的胞质。在细胞质中，这种DNA在感染后期没有转化为DNA酶抗性形式，这与未能从感染的细胞质中分离出物理颗粒相一致。用100 μ g利福平/ml从0-8小时处理牛痘病毒感染的细胞以增加病毒前体库，随后除去药物，导致病毒产量增加3倍。这种处理对病毒感染的细胞质没有影响。在一个单步生长周期的不同时间制备的病毒感染细胞和胞质体的薄切片中，通过EM研究了牛痘病毒的形态发生。虽然早期病毒形态发生形式出现，有没有随后的DNA凝聚或粒子成熟的细胞质。牛痘病毒可能需要宿主细胞核中的某些因子或功能才能正常成熟并产生感染性子代病毒。
Cytochalasin B-induced enucleation techniques were used to examine the ability of vaccinia virus to replicate in the absence of the host-cell nucleus in several mammalian cell lines. Virus-infected enucleated cells (cytoplasts) prepared from BSC-40 [African green monkey kidney], CVC and L [mouse fibroblast] cells were incapable of producing infectious progeny virus. The nature of this apparent nuclear involvement was studied in detail in BSC-40 cells. Modulations designed to maximize cytoplast integrity and longevity, such as reduction of the growth temperature and initial multiplicity of infection, did not improve virus growth in cytoplasts. Sodium dodecyl sulfate-polyacrylamide gel analysis of the [35S]methionine pulse-labeled proteins synthesized in vaccinia virus-infected cytoplasts demonstrated that early and late viral gene products were expressed at high levels and with the proper temporal sequence. Vaccinia virus cytoplasmic DNA synthesis, as measured by [3H]thymidine incorporation, peaked at 3 h postinfection and was 70-90% of control levels in cytoplasts. In the cytoplasts this DNA was not converted to a DNase-resistant form late in infection, which was consistent with the failure to isolate physical particles from infected cytoplasts. Treatment of vaccinia virus-infected cells with 100 .mu.g of rifampin/ml from 0-8 h to increase the pools of viral precursors, followed by subsequent removal of the drug, resulted in a 3-fold increase virus yield. This treatment had no effect on virus-infected cytoplasts. Vaccinia virus morphogenesis was studied by EM in thin sections of virus-infected cells and cytoplasts prepared at various times during a single-step growth cycle. Although early virus morphogenetic forms appeared, there was no subsequent DNA condensation or particle maturation in the cytoplasts. Vaccinia virus probably requires some factor or function from the host-cell nucleus in order to mature properly and produce infectious progeny virus.