Takagi T, Iwama M, Seta K, Kanda T, Tsukamoto T, Tominaga S, Mizumoto K
School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.
Arch Virol. 1996;141(9):1623-35. doi: 10.1007/BF01718287.
In vitro mRNA synthesis by Sendai virus is almost entirely dependent on the addition of cellular proteins (positive host factors), one of which could be tubulin. In this study, we investigated the distribution of host factors in various rat organs. Extracts from the brain, thymus, heart, lung, testis, ovary, and uterus all supported in vitro Sendai virus transcription, among which the highest activity was obtained with the brain extract. On the other hand, little or no activity was detected in the liver, spleen, and kidney extracts. An inverse correlation between the apparent host factor activity to stimulate mRNA synthesis and RNase activity that hydrolyzes Sendai virus mRNAs was found, except in the liver extract. However, when a transcription initiation complex was isolated and subjected to RNA chain elongation reaction, all of the extracts including those from liver, spleen and kidney, were active. Immunoblotting showed that tubulin molecules were integrated in these initiation complexes, supporting the notion that tubulin is involved in the initiation complex formation. We also identified a transcription inhibitory activity without any detectable RNase activity in the liver extract. This negative host factor seemed to act on RNA chain elongation. It is likely that Sendai virus transcription is regulated by both positive and negative regulatory factors.
仙台病毒在体外的mRNA合成几乎完全依赖于细胞蛋白(阳性宿主因子)的添加,其中之一可能是微管蛋白。在本研究中,我们调查了宿主因子在各种大鼠器官中的分布。来自脑、胸腺、心脏、肺、睾丸、卵巢和子宫的提取物均支持仙台病毒的体外转录,其中脑提取物的活性最高。另一方面,在肝脏、脾脏和肾脏提取物中几乎检测不到活性或无活性。除肝脏提取物外,发现刺激mRNA合成的表观宿主因子活性与水解仙台病毒mRNA的RNase活性之间呈负相关。然而,当分离转录起始复合物并进行RNA链延伸反应时,包括来自肝脏、脾脏和肾脏的提取物在内的所有提取物均具有活性。免疫印迹显示微管蛋白分子整合在这些起始复合物中,支持微管蛋白参与起始复合物形成的观点。我们还在肝脏提取物中鉴定出一种没有任何可检测到的RNase活性的转录抑制活性。这种阴性宿主因子似乎作用于RNA链延伸。仙台病毒转录很可能受正、负调控因子的共同调节。