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腺病毒12型基因组的转录。V. 增殖性感染中核RNA和信使RNA的动力学及大小分级分析。

Transcription of the genome of adenovirus type 12. V. Kinetic and size-class analyses of nuclear and messenger RNA in productive infection.

作者信息

Scheidtmann K H, Mansi-Wothke H, Doerfler W

出版信息

J Gen Virol. 1979 Sep;44(3):699-713. doi: 10.1099/0022-1317-44-3-699.

Abstract

In human KB cells productively infected with adenovirus type 12 (Ad12) the kinetics of synthesis of Ad12-specific RNA were investigated. Using 2 h labelling periods, striking differences were observed in the time course and the rate of synthesis of virus RNA comparing the nuclear, cytoplasmic and polysomal RNA fractions. A protracted onset of virus RNA synthesis was followed by an exponential increase between 15 and 40 h p.i.; reaching a maximum at 40 to 45 h p.i., the fraction of the newly synthesized Ad12-specific RNA remained constant up to 66 h in the nucleus, but decreased in the cytoplasm. Characteristic patterns were found in the size distribution of Ad12-specific messenger RNA synthesized early and late in the infection cycle. By using 10 min labelling periods, the size distribution and the degree of polyadenylation of the primary transcripts were determined. Early in infection, the size distribution of the virus-specific nuclear RNA resembled that of the mRNA, whereas late after infection most of the label was found in high mol. wt. RNA sedimenting between 30 and 55S. Eighty percent of the nuclear RNA was polyadenylated and belonged to the high mol. wt. RNA class. During the early phase of infection, approx. 20% of the virus genome was found to be transcribed symmetrically, and later after infection the entire Ad12 genome. Symmetrical (self-complementary) transcripts were preferentially drived from the terminal parts of Ad12 DNA.

摘要

在被12型腺病毒(Ad12)有效感染的人KB细胞中,研究了Ad12特异性RNA的合成动力学。使用2小时的标记期,比较细胞核、细胞质和多核糖体RNA组分时,在病毒RNA合成的时间进程和速率上观察到显著差异。病毒RNA合成开始延迟,随后在感染后15至40小时呈指数增长;在感染后40至45小时达到最大值,新合成的Ad12特异性RNA在细胞核中直至66小时保持恒定,但在细胞质中减少。在感染周期早期和晚期合成的Ad12特异性信使RNA的大小分布中发现了特征模式。通过使用10分钟的标记期,确定了初级转录本的大小分布和多聚腺苷酸化程度。感染早期,病毒特异性细胞核RNA的大小分布类似于mRNA,而感染后期大部分标记物存在于高分子量RNA中,沉降系数在30至55S之间。80%的细胞核RNA是多聚腺苷酸化的,属于高分子量RNA类别。在感染早期,约20%的病毒基因组被发现对称转录,感染后期则是整个Ad12基因组。对称(自我互补)转录本优先来自Ad12 DNA的末端部分。

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