The host Pex19p plays a role in peroxisomal localization of tombusvirus replication proteins
The host Pex19p plays a role in peroxisomal localization of tombusvirus replication proteins
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DOI：
10.1016/j.virol.2008.06.044
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发表时间：
2008-09-30
期刊：
影响因子：
3.7
通讯作者：
Nagy, Peter D.
中科院分区：
文献类型：
--
作者：
Pathak, Kunj B.;Sasvari, Zsuzsanna;Nagy, Peter D.
番茄丛矮病毒（TBSV）RNA的复制发生在植物和模式宿主酵母中过氧化物酶体的胞质膜表面。为了确定参与协助番茄丛矮病毒p33复制蛋白的过氧化物酶体定位的宿主蛋白，我们测试了p33是否可以直接结合到参与过氧化物酶体转运的酵母蛋白。这项工作已经通过下拉和共纯化实验证明了Pex 19 p-p33相互作用。Pex 19 p在番茄丛矮病毒复制酶蛋白交联后也被检测到，这表明Pex 19 p瞬时结合到复制酶上，这可能是预期的转运蛋白。为了验证Pex 19 p-p33相互作用在酵母中TBSV复制中的重要性，我们将Pex 19 p重新靶向线粒体，这导致大部分p33重新分布到线粒体。在体内，Pex 19 p的表达抑制TBSV RNA积累2-4倍，并降低Tombusvirus复制酶的体外活性80%。这些数据支持的模型，Pex 19 p是一个细胞转运蛋白的本地化的p33复制蛋白的宿主过氧化物酶体膜。(C)2008年爱思唯尔公司All rights reserved.
Replication of Tomato bushy stunt virus (TBSV) RNA takes place on the cytosolic membrane surface of peroxisomes in plants and in yeast, a model host. To identify the host proteins involved in assisting the peroxisomal localization of the tombusvirus p33 replication protein, we tested if p33 could bind directly to yeast proteins involved in peroxisomal transport in vitro. This work has led to the demonstration of Pex19p-p33 interaction via pull-down and co-purification experiments. Pex19p was also detected in the tombusvirus replicase after protein cross-linking, Suggesting that Pex19p transiently binds to the replicase as Could be expected from a transporter. To validate the importance of Pex19p-p33 interaction in TBSV replication in yeast, we re-targeted Pex19p to the mitochondria, which resulted in the re-distribution of a large fraction of p33 to the mitochondria. The expression of the mitochondrial-targeted Pex19p inhibited TBSV RNA accumulation by 2-4-fold in vivo and reduced the in vitro activity of the tombusvirus replicase by 80%. These data Support the model that Pex19p is a cellular transporter for localization of p33 replication protein to the host peroxisomal membranes. (C) 2008 Elsevier Inc. All rights reserved.