CHARACTERIZATION OF MESSENGER-RNA OF INFLUENZA-VIRUS
CHARACTERIZATION OF MESSENGER-RNA OF INFLUENZA-VIRUS
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DOI：
10.1128/jvi.16.6.1435-1443.1975
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发表时间：
1975-01-01
影响因子：
5.4
通讯作者：
BARRY, RD
中科院分区：
文献类型：
--
作者：
GLASS, SE;MCGEOCH, D;BARRY, RD
流感病毒mRNA出现的动力学、mRNA在游离和膜相关多聚核糖体之间的分布、其poly（A）含量以及基因组在感染早期转录成mRNA的程度被确定。从流感病毒感染的细胞中制备多聚核糖体，在感染后的不同时间用[3 H]尿苷标记30分钟。从这些多聚核糖体中提取的大多数3 H标记的RNA在蔗糖梯度中沉淀为不均匀的8 S至20 S峰，并且它在很大程度上与病毒体RNA互补。根据以下标准，互补RNA具有通常归因于mRNA的性质：（i）它被[3 H]尿苷快速标记;（ii）在戊二醛处理后，它在CsCl密度梯度中与多聚核糖体结合;（iii）它含有聚（A）。在37 ℃的鸡细胞中，病毒mRNA在感染后45分钟开始出现，在感染后2 ~ 2.5小时出现最多。分离感染细胞的游离和膜结合的多聚核糖体，发现它们含有相同种类的mRNA。有没有绝对的分离的mRNA序列到任何多聚核糖体类，虽然每个可能包含不同的mRNA的不同比例。从感染后45分钟开始，膜结合和游离的多聚核糖体含有poly（A）的RNA含有至少80%的基因组RNA互补的序列，而poly（A）-负RNA含有90至100%的基因组互补的序列。没有证据表明流感病毒mRNA转录的时间控制。在31 ℃，当病毒发展相对于37 ℃减慢时，互补RNA在感染后1小时首次出现。此时，总多核糖体RNA含有与整个基因组互补的序列。
The kinetics of the appearance of influenza mRNA, the distribution of mRNA between free and membrane-associated polyribosomes, its poly(A) content, and the extent to which the genome was transcribed into mRNA early in infection were determined. Polyribosomes were prepared from influenza virus-infected cells labeled for 30-min periods at various times after infection with [3H]uridine. Most of the 3H-labeled RNA extracted from these polyribosomes sedimented as a heterogeneous 8S to 20S peak in sucrose gradients, and it was largely complementary to virion RNA. By the following criteria, the complementary RNA had properties normally ascribed to mRNA: (i) it labeled rapidly with [3H]uridine; (ii) after glutaraldelyde treatment, it banded with polyribosomes in CsCl density gradients; and (iii) it contained poly(A). In chick cells at 37 C, virus mRNA was first detectable at 45 min postinfection and reached its maximal rate of appearance at 2 to 2.5 h postinfection. The free and membrane-bound polyribosomes of infected cells were separated and were found to contain the same classes of mRNA. There was no absolute segregation of mRNA sequences into either polyribosome class although each probably contained distinct ratios of the different mRNA's. From 45 min postinfection onwards, both membrane-bound and free polysomal poly(A)-containing RNA contained sequences complementary to at least 80% of the genome RNA, whereas poly(A)-minus RNA contained sequences complementary to 90 to 100% of the genome. There was no evidence for the temporal control of transcription of influenza mRNA. At 31 C, when virus development was slowed relative to 37 C,complementary RNA first appeared at 1 h postinfection. At this time, total polysomal RNA contained sequences complementary to the whole genome.