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[感染口蹄疫病毒的细胞中10S颗粒的合成]

[Synthesis of 10S particles in cells infected with aphthovirus].

作者信息

Gómez Yafal A, Polacino P, Kaplan G, Palma E L

机构信息

Departamento de Virología, Centro de Investigaciones en Ciencias Veterinarias, I.N.T.A., Buenos Aires, Argentina.

出版信息

Rev Argent Microbiol. 1984;16(2):87-92.

PMID:6101037
Abstract

In the present study, evidence is presented for the existence of a morphogenetic intermediary that may be a precursor of the procapsids in the assembling process. BHK21 clone 13S cells were infected with Aphthovirus A24 (Cruzeiro strain), and pulse-chase experiments were carried out using 3H-leucine. Cytoplasmic extracts were then prepared at appropriate times, and analyzed by sucrose-gradient ultracentrifugation. After preliminary assays (Fig. 1), working conditions were standardized so as to obtain maximal recovery of the morphogenetic intermediary, as well as consistency of results. Only in the presence of DOC-Brij58 and Mg++ could a 10S sedimentation coefficient peak be seen (Fig. 1 a). A heterogeneous zone, with 4,5-5S as sedimentation coefficient, was also observed. The degree of labeling in the region 4,5-5S compared with that in the 10S portion, depends on the time within the infectious cycle when cells were pulse-labeled. Maximal levels for the ratio 10S/4,5-5S are reached when pulse-labeling takes place at the time when the amount of RNA viral synthesis reaches 80% of its total value (Fig. 2). Similar experiments, performed with third passage bovine fetal kidney cells, were confirmatory of the presence of a structure sedimenting at 10S, as well as of a heterogeneous zone of 4,5-5S (Fig. 3). It would appear that assembling of Aphthoviruses is accomplished through an intermediary unit which differs from that found for other Picornaviruses, the latter being the result of the union of 12 pentamers. The capsid of Aphthoviruses, also composed of 60 identical sub-units, would instead derive from the joining of 20 trimers.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

在本研究中,有证据表明存在一种形态发生中间体,它可能是装配过程中前衣壳的前体。用口蹄疫病毒A24(克鲁塞罗毒株)感染BHK21克隆13S细胞,并使用³H-亮氨酸进行脉冲追踪实验。然后在适当的时间制备细胞质提取物,并通过蔗糖梯度超速离心进行分析。经过初步测定(图1),对实验条件进行了标准化,以实现形态发生中间体的最大回收率以及结果的一致性。只有在存在脱氧胆酸钠-聚山梨醇酯58(DOC-Brij58)和镁离子的情况下,才能看到一个沉降系数为10S的峰(图1a)。还观察到一个沉降系数为4.5-5S的异质区。4.5-5S区域与10S部分的标记程度取决于细胞进行脉冲标记时在感染周期内的时间。当在RNA病毒合成量达到其总量的80%时进行脉冲标记时,10S/4.5-5S的比例达到最高水平(图2)。用第三代牛胎儿肾细胞进行的类似实验证实了存在沉降系数为10S的结构以及4.5-5S的异质区(图3)。看来口蹄疫病毒的装配是通过一个不同于其他小RNA病毒的中间单元完成的,后者是12个五聚体结合的结果。口蹄疫病毒的衣壳同样由60个相同的亚基组成,相反,它将来自20个三聚体的连接。(摘要截断于250字)

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