Localization of the 26-S RNA sequence on the viral genome type 42-S RNA isolated from SFV-infected cells.
Localization of the 26-S RNA sequence on the viral genome type 42-S RNA isolated from SFV-infected cells.
复制标题
从 SFV 感染细胞中分离出的病毒基因组 42-S RNA 类型中 26-S RNA 序列的定位。
DOI：
10.1016/0042-6822(76)90073-8
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发表时间：
1976
期刊：
影响因子：
3.7
通讯作者：
G. Wengler
中科院分区：
文献类型：
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作者：
G. Wengler
32P 标记的病毒特异性 42- 和 26-S RNA 是从感染披膜病毒 Semliki 森林病毒的 BHK 21 细胞中制备的。用 RNAse T1 消化后，通过二维聚丙烯酰胺凝胶电泳（T1 指纹）对这些 RNA 种类产生的寡核苷酸进行分析，结果表明这两种 RNA 种类均含有 Poly(A) 序列，并且 26-S RNA 的寡核苷酸代表 42-S RNA 中存在的寡核苷酸的子集。以下发现表明，42-S RNA 上存在的不同核苷酸序列的顺序为 5'-末端-42-S RNA 特异性序列-26-S RNA 序列-poly(A)-AOH。 (1) 在受感染细胞中积累的 26-S RNA 和在蔗糖密度梯度上沉积在 26 S 周围的 42-S RNA 含 Poly(A) 片段的 T1 指纹中发现相同的寡核苷酸模式。 (2)从42-和26-S RNA的T1指纹中分离出poly(A)片段，用胰腺RNA酶消化，并在DEAE纸上通过电泳分析反应产物。结果表明，两个片段均含有由U6C2(AC)1(AU)1(AAU)1组成的杂聚序列。 (3)到目前为止，除了含poly(A)的寡核苷酸外，在所有分析过的T1指纹来源的寡核苷酸中均检测到了GMP，表明后者含有其来源分子的3'末端。 (4)用RNAse T1和胰腺RNAse消化[3H]腺苷标记的42-S RNA，分离出poly(A)片段，并测定这些片段的[3H]腺苷与[3H]AMP放射性比为1:97。讨论了 42-S RNA 上不同序列的组织对其翻译和 SFV 复制的可能影响。
32P-labeled virus-specific 42- and 26-S RNA has been prepared from BHK 21 cells infected with the togavirus Semliki Forest virus. Analyses of the oligonucleotides generated from these RNA species after digestion with RNAse T1by two-dimensional polyacrylamide gel electrophoresis (T1-fingerprints) show that both RNA species contain poly(A) sequences and that the oligonucleotides of the 26-S RNA represent a subset of those present in 42-S RNA. The following findings indicate that the different nucleotide sequences are present on the 42-S RNA in the order 5′-terminus-42-S RNA-specific sequences-26-S RNA sequences-poly(A)-AOH. (1) Identical oligonucleotide patterns are found in the T1-fingerprints of the 26-S RNA accumulating in infected cells and of poly(A)-containing fragments of the 42-S RNA which sediment around 26 S on sucrose density gradients. (2) The poly(A) fragments were isolated from T1-fingerprints of 42-and 26-S RNA, digested with pancreatic RNAse, and the reaction products analyzed by electrophoresis on DEAE paper. The results indicate that both fragments contain a heteropolymeric sequence consisting of U6C2(AC)1(AU)1(AAU)1. (3) GMP has been detected in all oligonucleotides derived from T1-fingerprints which have been analyzed so far, except in the poly(A)-containing oligonucleotides, indicating that the latter contain the 3′-termini of the molecules from which they were derived. (4) [3H]Adenosine-labeled 42-S RNA was digested with RNAse T1and pancreatic RNAse, the poly(A) fragments were isolated, and an [3H]adenosine to [3H]AMP radioactivity ratio of 1 to 97 was determined for these fragments. Possible implications of the organization of the different sequences on the 42-S RNA on its translation and the replication of SFV are discussed.