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[甲醛诱导蛋白质与杆状病毒颗粒中的RNA紧密结合]

[Formaldehyde-induced tight binding of protein to RNA in particles of rod-like virus].

作者信息

Mazhul' L A, Dobrov E N, Tikhonenko T I

出版信息

Biokhimiia. 1978 Jan;43(1):138-45.

PMID:564215
Abstract

The formaldehyde-induced formation of tightly bound RNA-protein complexes of rod-like plant viruses was studied. The preparations of tobacco mosaic virus and closely related cucumber virus 4 were incubated with 1.5% formaldehyde for 20-50 hrs at 50 degrees C. Then the viral particles were disrupted, free protein was removed and viral RNA was centrifuged in the linear gradient of Cs2SO4. The RNAs from the formaldehyde-untreated viruses and RNA from the formaldehyde-treated tobacco masaic virus had the density of 1.65-1.66 g/cm3, while RNA from the formaldehyde-treated cucumber virus had the density of 1.57-1.42 g/cm3, depending on the incubation time. This is indicative of the protein binding to RNA. Treatment of the cucumber virus complex with pronase resulted in a liberation of free RNA with the density of 1.66 g/cm3; incubation for 2 min at 100 degrees C in a dissociating mixture (2% sodium dodecyl sulfate + 0.2% mercaptoethanol) did not cause the dissociation of the complex. Polyacrylamide gel electrophoresis showed that the most part of the protein molecules are bound within the complex not by covalent protein-protein cross-links.

摘要

研究了甲醛诱导杆状植物病毒形成紧密结合的RNA-蛋白质复合物的过程。将烟草花叶病毒制剂和密切相关的黄瓜病毒4制剂在50℃下与1.5%甲醛孵育20至50小时。然后破坏病毒颗粒,去除游离蛋白质,并将病毒RNA在硫酸铯线性梯度中离心。未用甲醛处理的病毒的RNA以及用甲醛处理的烟草花叶病毒的RNA密度为1.65 - 1.66 g/cm³,而用甲醛处理的黄瓜病毒的RNA密度为1.57 - 1.42 g/cm³,这取决于孵育时间。这表明蛋白质与RNA结合。用链霉蛋白酶处理黄瓜病毒复合物会释放出密度为1.66 g/cm³的游离RNA;在解离混合物(2%十二烷基硫酸钠 + 0.2%巯基乙醇)中于100℃孵育2分钟不会导致复合物解离。聚丙烯酰胺凝胶电泳表明,大部分蛋白质分子在复合物中并非通过共价蛋白质-蛋白质交联结合。

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