Cowpea mosaic virus 32- and 60-kilodalton replication proteins target and change the morphology of endoplasmic reticulum membranes
Cowpea mosaic virus 32- and 60-kilodalton replication proteins target and change the morphology of endoplasmic reticulum membranes
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DOI：
10.1128/jvi.76.12.6293-6301.2002
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发表时间：
2002-06-01
影响因子：
5.4
通讯作者：
van Kammen, A
中科院分区：
文献类型：
--
作者：
Carette, JE;van Lent, J;van Kammen, A
豇豆花叶病毒（CPMV）的复制与细胞内膜重排形成的小膜泡密切相关。为了确定哪些病毒蛋白负责膜的重排和复制复合物的附着，我们已经表达了由RNA 1编码的个体CPMV蛋白在豇豆原生质体中通过瞬时表达和在本氏烟草植物中通过使用烟草脆裂病毒（TRV）表达载体。当单独表达时，32-kDa蛋白（32 K）和60 K仅以少量积累，但发现与主要来自内质网（ER）的膜相关。24 K和110 K是易溶的，并积累到高水平。使用TRV载体，32 K和60 K的表达导致ER膜的重排。此外，32 K和60 K的表达导致接种N.本塞姆氏藻叶片中的32 K和60 K蛋白，表明32 K和60 K是细胞毒蛋白。另一方面，在CPMV感染期间，32 K和60 K积累到高水平而不引起坏死。
Cowpea mosaic virus (CPMV) replicates in close association with small membranous vesicles that are formed by rearrangements of intracellular membranes. To determine which of the viral proteins are responsible for the rearrangements of membranes and the attachment of the replication complex, we have expressed individual CPMV proteins encoded by RNA1 in cowpea protoplasts by transient expression and in Nicotiana benthamiana plants by using the tobacco rattle virus (TRV) expression vector. The 32-kDa protein (32K) and 60K, when expressed individually, accumulate in only low amounts but are found associated with membranes mainly derived from the endoplasmic reticulum (ER). 24K and 110K are freely soluble and accumulate to high levels. With the TRV vector, expression of 32K and 60K results in rearrangement of ER membranes. Besides, expression of 32K and 60K results in necrosis of the inoculated N. benthamiana leaves, suggesting that 32K and 60K are cytotoxic proteins. On the other hand, during CPMV infection 32K and 60K accumulate to high levels without causing necrosis.