Structure of the pseudorabies virus capsid: comparison with herpes simplex virus type 1 and differential binding of essential minor proteins.
Structure of the pseudorabies virus capsid: comparison with herpes simplex virus type 1 and differential binding of essential minor proteins.
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DOI：
10.1016/j.jmb.2013.06.034
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发表时间：
2013-09-23
影响因子：
5.6
通讯作者：
Conway, J. F.
中科院分区：
文献类型：
--
作者：
Homa, F. L.;Huffman, J. B.;Toropova, K.;Lopez, H. R.;Makhov, A. M.;Conway, J. F.
用冷冻电镜（cryo-EM）观察了从感染细胞核和伪狂犬病毒（PRV）病毒粒子中分离的PRV衣壳的结构，并与单纯疱疹病毒1型（HSV-1）衣壳进行了比较。PRV衣壳结构与HSV-1的衣壳结构非常相似，包括HSV-1的衣壳顶点特异性组分（CVSC）的分布，CVSC是pUL 17和pUL 25蛋白的异源二聚体。CVSC在所有PRV衣壳上的占有率接近100%，相比之下，对于HSV-1 C-衣壳报道的占有率为约50%，对于HSV-1 A-和B-衣壳我们测量的占有率为25%或更低。缺乏pUL 25的PRV突变体不产生C-衣壳，并且在基于冷冻EM的重建中缺乏可见的CVSC密度。其中绿色荧光蛋白（GFP）融合在pUL 25的N-末端内的PRV衣壳的重建证实了先前的研究，其中类似的HSV-1衣壳突变体将pUL 25定位于五邻体远端的CVSC密度区。然而，在9 μ ngstrom分辨率PRV C-衣壳图谱中的CVSC密度与HSV-1 pUL 25的可用晶体结构的比较未能找到令人满意的拟合，这表明PRV pUL 25的不同折叠或pUL 25的衣壳结合构象与从溶液中结晶的蛋白质确定的X射线模型不匹配。在病毒粒子内成像的PRV衣壳非常类似于C-衣壳，其中增加了弱但显著的密度，包围了我们归因于被膜蛋白的五邻体。我们的研究结果表明PRV和HSV衣壳之间具有显著的结构保守性。
The structure of pseudorabies virus (PRV) capsids isolated from the nucleus of infected cells and from PRV virions was determined by cryo-electron microscopy (cryo-EM), and compared to herpes simplex virus type 1 (HSV-1) capsids. PRV capsid structures closely resemble those of HSV-1, including distribution of the capsid vertex specific component (CVSC) of HSV-1, which is a heterodimer of the pUL17 and pUL25 proteins. Occupancy of CVSC on all PRV capsids is near 100%, compared to ~50% reported for HSV-1 C-capsids and 25% or less that we measure for HSV-1 A- and B-capsids. A PRV mutant lacking pUL25 does not produce C-capsids and lacks visible CVSC density in the cryo-EM-based reconstruction. A reconstruction of PRV capsids in which green fluorescent protein (GFP) was fused within the N-terminus of pUL25 confirmed previous studies with a similar HSV-1 capsid mutant localizing pUL25 to the CVSC density region that is distal to the penton. However, comparison of the CVSC density in a 9 Ångstrom resolution PRV C-capsid map with the available crystal structure of HSV-1 pUL25 failed to find a satisfactory fit, suggesting either a different fold for PRV pUL25 or a capsid- bound conformation for pUL25 that does not match the X-ray model determined from protein crystallized in solution. The PRV capsid imaged within virions closely resembles C-capsids with the addition of weak but significant density shrouding the pentons that we attribute to tegument proteins. Our results demonstrate significant structure conservation between the PRV and HSV capsids.
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影响因子：
5.6
作者：
Conway JF;Cockrell SK;Copeland AM;Newcomb WW;Brown JC;Homa FL
通讯作者：
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