Anatomy of herpes simplex virus DNA. III. Characterization of defective DNA molecules and biological properties of virus populations containing them
Anatomy of herpes simplex virus DNA. III. Characterization of defective DNA molecules and biological properties of virus populations containing them
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单纯疱疹病毒 DNA 的解剖。
作者：
Niza Frenkel;Robert J. Jacob;R. Honess;Gary S. Hayward;Hilla Locker;Bernard Roizman
我们在Hep-2细胞中鉴定了单纯疱疹病毒1型空斑纯化和未稀释传代的病毒后代及其DNA。感染病毒产量在第1~10代呈递减趋势，在第11~14代呈逐渐上升趋势，与PFU/颗粒比相关。(Ii)在感染第6代至第10代病毒的细胞中，早期病毒多肽(4号)产生过剩，晚期病毒蛋白的合成延迟。此外，这些传代中的病毒干扰了非缺陷标记病毒的复制。感染第14代病毒的细胞产生正常数量的多肽4，而且，该病毒显示出最小的干扰能力。(3)除了密度为1.726 g/cm-3的脱氧核糖核酸是第0代病毒后代中存在的唯一成分外，第6代至第14代还含有一个额外的物种(p 1.732)，在某些情况下(第6代和第10代)还含有中等浮力密度的脱氧核糖核酸。P 1.732与p 1.726 DNA的比例在第6~9代中增加到最大值4，在第10~14代中逐渐下降到1。(4)p 1.732 DNA与p 1.726 DNA在大小上不能区分，但它没有HinⅢ酶切位点，经EcoRI酶消化后只产生两种相对分子质量分别为5.1x10-6和5.4x10-6的片段。(V)从第14代纯化的p 1.732 DNA的部分变性图谱显示，存在两种类型的重复单位，其大小大致与EcoRI片段相同，位于不同的分子中。(Vi)除了由不同大小的串联修复单元组成的两种p-1.732分子外，通过比较不同代次的后代的特定感染力和干扰能力，还出现了缺陷dna分子多样性的其他证据。数据表明，一些带有正常浮力密度(1.726)的dna的颗粒也一定是有缺陷的，因为干扰和产生过量多肽4的能力似乎与高浮力密度有缺陷的dna的数量不成正比。这些数据表明，有缺陷的干扰颗粒被干扰能力减弱的有缺陷的颗粒所取代，并且在HSV的连续制备过程中进化出不止一种有缺陷的DNA分子。
We have characterized the virus progeny and its DNA from plaque-purified and undiluted passages of herpes simplex virus 1 in HEp-2 cells. Secifically, (i) infectious virus yields declined progressively in passages 1 through 10 and gradually increased at passages 11 through 14. The yields correlated with PFU/particle ratios. (ii) In cells infected with virus from passages 6 through 10, there was an overproduction of an early viral polypeptide (no. 4) and a delay in the synthesis of late viral proteins. In addition, the virus in these passages interfered with the replication of a nondefective marker virus. Cells infected with passage 14 virus produced normal amounts of polypeptide 4 and, moreover, this virus showed minimal interfering capacity. (iii) In addition to DNA of density 1.726 g/cm-3, which was the sole component present in viral progeny of passage 0, passages 6 through 14 contained one additional species (p 1.732) and in some instances (passages 6 and 10) also DNA of an intermediate buoyant density. The ratio of p 1.732 to p 1.726 DNA increased to a maximum of 4 in passages 6 through 9 and gradually decreased to 1 in passages 10 through 14. (iv) p 1.732 DNA cannot be differentiated from p 1.726 DNA with respect to size; however, it has no Hin III restriction enzyme cleavage sites and yields only predominantly two kinds of fragments with molecular weights of 5.1 x 10-6 and 5.4 x 10-6 upon digestion with EcoRI enzyme. (v) Partial denaturation profiles of purified p 1.732 DNA from passage 14 revealed the presence of two types of tandemly repeated units corresponding roughly in size to the EcoRI fragments and situated in different molecules. (vi) In addition to the two kinds of p 1.732 molecules consisting of tandem repaeat units of different sizes, other evidence for the diversity of defective DNA molecules emerged from comparisons of specific infectivity and interfering capacity of the progeny from various passages. The data suggest that some of the particles with DNA of normal buoyant density (1.726) must also be defective since the capacity to interfere and to produce an excess of polypeptide 4 did not appear to be proportional to the amount of high-buoyant-density defective DNA. The data suggest that defective interfering particles are replaced by defective particles with diminished capacity to interfere and that more than one species of defective DNA molecules evolves on serial preparation of HSV.