Molecular characterization of tomato spotted Wilt virus defective interfering RNAs and detection of truncated L proteins.
Molecular characterization of tomato spotted Wilt virus defective interfering RNAs and detection of truncated L proteins.
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番茄斑枯萎病病毒缺陷干扰 RNA 的分子表征和截短 L 蛋白的检测。
作者：
A. Inoue;R. Kormelink;J. Sgro;Tatsuya Nagata;E. W. Kitajima;R. Goldbach;Dick Peters
对番茄斑萎病毒（TSWV）25个不同的缺陷型干扰RNA（DI RNA）的连接位点进行了分析。DI RNA的大小从2.0到5.2个碱基（kb）变化，并且含有单个内部缺失。小于2kb的DI RNA的缺乏表明TSWV DI RNA的存活的大小限制。这一假设得到了由两个1.6长的单体首尾相连形成的二聚体DI RNA的发现的加强。发现了三种类型的连接位点，一种类型起源于简单的缺失;第二种类型含有一些来源不明的额外核苷酸;第三种类型含有三到五个核苷酸的延伸，最初发生在缺失的两侧，其中一个被删除。在研究的25个DI RNA中，有19个保持了原始的阅读框架，表明具有翻译效力的DI RNA的选择性偏好。在核衣壳制备物中确实检测到由这些DI RNA编码的截短蛋白。完整的L RNA的折叠研究表明，计算的最小折叠能量在16 ℃时低于23 ℃时，表明该分子在低温下具有更高的稳定性。结果表明，在缺失的过程中，局部折叠的二级结构的参与，而不是在缺失点周围的某些序列的要求。如所讨论的，TSWV中DI RNA的产生基本上类似于许多病毒中描述的RNA重组过程。
Junction sites of 25 different defective interfering (DI) RNAs of tomato spotted wilt virus (TSWV) were characterized. The DI RNAs varied in size from 2.0 to 5.2 kilobases (kb) and contained a single internal deletion. The absence of DI RNAs smaller than 2 kb suggested a size constraint for the survival of TSWV DI RNAs. This hypothesis was reinforced by the finding of a dimeric DI RNA formed by two 1.6-long monomers linked head to tail. Three types of junction sites were found, one type originating from a simple deletion; the second type contained a few extra nucleotides of unknown origin; and the third type contained a stretch of three to five nucleotides, originally occurring at both sides of the deletion and of which one was deleted. In 19 of the 25 DI RNAs studied, the original reading frame was maintained, suggesting a selective preference of DI RNAs with translational potency. Truncated proteins encoded by these DI RNAs were indeed detected in the nucleocapsid preparations. Folding studies of the complete L RNA revealed that the calculated minimal energy of folding was at 16 degreesC lower than at 23 degrees, indicating a higher stability of this molecule at low temperatures. The results suggest an involvement of locally folded secondary structures in the process of deletion, rather than the requirement of certain sequences around the deletion point. The DI RNA generation in TSWV is essentially, as discussed, similar to the process of RNA recombination described in many viruses.
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DOI：
10.1006/viro.1995.1515
发表时间：
1995
期刊：
Virology.
影响因子：
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作者：
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10.1073/pnas.86.20.7706
发表时间：
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影响因子：
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作者：
JAEGER, JA;TURNER, DH;ZUKER, M
通讯作者：
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DOI：
10.1006/viro.1995.1163
发表时间：
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期刊：
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影响因子：
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DOI：
10.1006/viro.1996.8377
发表时间：
1997
期刊：
Virology.
影响因子：
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作者：
Pogany,J;Romero,J;Bujarski,JJ
通讯作者：
Bujarski,JJ