Analysis of potato virus X replicase and TGBp3 subcellular locations
Analysis of potato virus X replicase and TGBp3 subcellular locations
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DOI：
10.1016/j.virol.2009.08.002
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发表时间：
2009-10-25
期刊：
影响因子：
3.7
通讯作者：
Verchot-Lubicz, Jeanmarie
中科院分区：
文献类型：
--
作者：
Bamunusinghe, Devinka;Hemenway, Cynthia L.;Verchot-Lubicz, Jeanmarie
马铃薯X病毒(PVX)侵染引起寄主内膜系统的某些细胞病理改变。PVX复制酶的亚细胞位置以前是未知的，而PVX TGBp3蛋白先前被报道驻留在内质网。利用表达绿色荧光蛋白报告基因的PVX感染性克隆，以及检测PVX复制酶和宿主膜标志物的抗血清，我们检测了PVX复制酶与TGBp3的亚细胞分布。共聚焦和电子显微镜观察表明，复制酶定位于内质网衍生的膜结合结构中。TGBp3的一个子集驻留在与复制酶相同位置的内质网中。蔗糖梯度分级显示PVX复制酶和TGBp3蛋白与ER标志蛋白共分级。这种定位代表了两种蛋白质都可以合成和/或发挥功能的区域。没有证据表明PVX蛋白进入高尔基体。抑制新生膜合成的药物Cerulenin也抑制了PVX的复制。这些综合数据表明，PVX的复制依赖于内质网衍生的膜募集和膜增殖。由爱思唯尔公司出版。
Potato virus X (PVX) infection leads to certain cytopathological modifications of the host endomembrane system. The subcellular location of the PVX replicase was previously unknown while the PVX TGBp3 protein was previously reported to reside in the ER. Using PVX infectious clones expressing the green fluorescent protein reporter, and antisera detecting the PVX replicase and host membrane markers, we examined the subcellular distribution of the PVX replicase in relation to the TGBp3. Confocal and electron microscopic observations revealed that the replicase localizes in membrane bound structures that derive from the ER. A subset of TGBp3 resides in the ER at the same location as the replicase. Sucrose gradient fractionation showed that the PVX replicase and TGBp3 proteins co-fractionate with ER marker proteins. This localization represents a region where both proteins may be synthesized and/or function. There is no evidence to indicate that either PVX Protein moves into the Golgi apparatus. Cerulenin, a drug that inhibits de novo membrane synthesis, also inhibited PVX replication. These combined data indicate that PVX replication relies on ER-derived membrane recruitment and membrane proliferation. Published by Elsevier Inc.