Lysine 242 within Helix 10 of the Pseudorabies Virus Nuclear Egress Complex pUL31 Component Is Critical for Primary Envelopment of Nucleocapsids
Lysine 242 within Helix 10 of the Pseudorabies Virus Nuclear Egress Complex pUL31 Component Is Critical for Primary Envelopment of Nucleocapsids
复制标题
伪狂犬病病毒核出口复合物 pUL31 成分螺旋 10 内的赖氨酸 242 对于核衣壳的初级包膜至关重要
DOI：
10.1128/jvi.01182-17
复制
发表时间：
2017
影响因子：
5.4
通讯作者：
Mettenleiter TC
中科院分区：
文献类型：
--
作者：
Rönfeldt S;Klupp BG;Franzke K;Mettenleiter TC
新组装的疱疹病毒核衣壳通过囊泡介导的过程与核膜接合，从细胞核转移到细胞质。这种转运是由保守的核出口复合体（NEC）控制的，由字母疱疹病毒pUL34和pUL31同源物组成。NEC不仅是有效的核出口所必需的，而且也足以从内核膜（INM）形成囊泡，以及合成脂质双分子层。最近解决的来自不同疱疹病毒的NECs晶体结构揭示了这种膜变形和断裂机制的分子细节，揭示了参与复合物和外壳形成的界面。然而，与核衣壳的相互作用域仍未确定。由于NEC在INM的核质侧组装了一个弯曲的六角形外壳，该外壳由紧密交织的pUL31/pUL34异源二聚体组成，排列在六聚体中，因此只有由pUL31残基形成的NEC的膜远端才能与核衣壳cargo相互作用。为了鉴定参与衣壳结合的氨基酸，我们对甲型疱疹病毒伪狂犬病毒（PrV）的相应区域进行了突变。我们检测了位点特异性突变的pUL31同源物的定位、与pUL34的相互作用以及PrV-ΔUL31的互补。我们在PrV pUL31中发现了一个保守的赖氨酸残基，它位于NEC膜远端暴露的α -螺旋结构域H10的242个氨基酸位置，是核衣壳融入新生初级颗粒的关键残基。通过核膜的囊泡运输是研究的焦点，但仍未得到很好的理解。疱疹病毒开创了这种机制，将新组装的核衣壳通过来自内膜的囊泡从细胞核转运到细胞质中，这些囊泡与外核膜融合在一个良好的调节过程中释放它们的内容物。病毒核膜出芽和断裂机制的结构最近得到了解决，提供了深入的分子细节。然而，货物是如何合并的仍不清楚。我们在核出口复合体的膜远端部分发现了一个保守的赖氨酸残基，这是衣壳摄取到核膜衍生的内囊泡所必需的。
Newly assembled herpesvirus nucleocapsids are translocated from the nucleus to the cytosol by a vesicle-mediated process engaging the nuclear membranes. This transport is governed by the conserved nuclear egress complex (NEC), consisting of the alphaherpesviral pUL34 and pUL31 homologs. The NEC is not only required for efficient nuclear egress but also sufficient for vesicle formation from the inner nuclear membrane (INM), as well as from synthetic lipid bilayers. The recently solved crystal structures for the NECs from different herpesviruses revealed molecular details of this membrane deformation and scission machinery uncovering the interfaces involved in complex and coat formation. However, the interaction domain with the nucleocapsid remained undefined. Since the NEC assembles a curved hexagonal coat on the nucleoplasmic side of the INM consisting of tightly interwoven pUL31/pUL34 heterodimers arranged in hexamers, only the membrane-distal end of the NEC formed by pUL31 residues appears to be accessible for interaction with the nucleocapsid cargo. To identify the amino acids involved in capsid incorporation, we mutated the corresponding regions in the alphaherpesvirus pseudorabies virus (PrV). Site-specifically mutated pUL31 homologs were tested for localization, interaction with pUL34, and complementation of PrV-ΔUL31. We identified a conserved lysine residue at amino acid position 242 in PrV pUL31 located in the alpha-helical domain H10 exposed on the membrane-distal end of the NEC as a key residue for nucleocapsid incorporation into the nascent primary particle.IMPORTANCEVesicular transport through the nuclear envelope is a focus of research but is still not well understood. Herpesviruses pioneered this mechanism for translocation of the newly assembled nucleocapsid from the nucleus into the cytosol via vesicles derived from the inner nuclear membrane which fuse in a well-tuned process with the outer nuclear membrane to release their content. The structure of the viral nuclear membrane budding and scission machinery has been solved recently, providing in-depth molecular details. However, how cargo is incorporated remained unclear. We identified a conserved lysine residue in the membrane-distal portion of the nuclear egress complex required for capsid uptake into inner nuclear membrane-derived vesicles.
登录
查看更多内容
影响因子：
6.7
作者：
Funk C;Ott M;Raschbichler V;Nagel CH;Binz A;Sodeik B;Bauerfeind R;Bailer SM
通讯作者：
Bailer SM
影响因子：
4.8
作者：
Lorenz, Michael;Vollmer, Benjamin;Antonin, Wolfram
通讯作者：
Antonin, Wolfram
影响因子：
16.6
作者：
Bigalke, Janna M.;Heuser, Thomas;Nicastro, Daniela;Heldwein, Ekaterina E.
通讯作者：
Heldwein, Ekaterina E.
影响因子：
5.4
作者：
Klupp, Barbara G.;Granzow, Harald;Mettenleiter, Thomas C.
通讯作者：
Mettenleiter, Thomas C.
DOI：
10.1073/pnas.97.20.11002
发表时间：
2000-09
影响因子：
11.1
作者：
G. Ye;B. Roizman
通讯作者：
G. Ye;B. Roizman