Infectious bursal disease virus VP5 polypeptide: a phosphoinositide-binding protein required for efficient cell-to-cell virus dissemination.
Infectious bursal disease virus VP5 polypeptide: a phosphoinositide-binding protein required for efficient cell-to-cell virus dissemination.
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DOI：
10.1371/journal.pone.0123470
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Rodríguez JF
中科院分区：
文献类型：
--
作者：
Méndez F;de Garay T;Rodríguez D;Rodríguez JF
传染性法氏囊病病毒(IBDV)属于禽类病毒科，是雏鸡免疫抑制病的主要病原体。IBDV基因组由两个dsRNA片段组成。最大的一个含有两个部分重叠的开放阅读框，分别编码一种被称为VP5的非结构多肽和一个大的多蛋白。VP5对病毒复制不是必需的。然而，它在IBDV的致病过程中起着重要作用。VP5积聚在IBDV感染细胞的质膜上。我们已经分析了VP5 PM目标的机制。更新的拓扑预测算法服务器无法识别VP5序列内的跨膜结构域。然而，VP5的多阳离子C-末端区域含有三个由两到三个连续的碱性氨基酸残基(赖氨酸或精氨酸)形成的紧密相连的斑块，这可能是其PM取向的原因。我们发现，无论是C-端VP5缺失还是碱性氨基酸被丙氨酸残基取代，减少VP5 C-端的正电荷的突变都会取消PM靶向。用亲和纯化的Flag标记的VP5(FVP5)蛋白版本进行的脂质重叠分析表明，该多肽与几个磷脂酰肌醇(PIP)结合，显示出明显的对单磷酸物种的偏好。用缺乏聚阳离子结构域的FVP5突变蛋白进行的实验表明，该区域对于PIP结合是必不可少的。用反向遗传学产生的表达C末端缺失的VP5多肽的IBDV突变体收集的数据表明，VP5-PIP结合域对于其在感染细胞中的PM靶向和有效的病毒传播都是必需的。这里提供的数据使我们假设IBDV可能使用一种非裂解的VP5依赖的细胞间传播机制。
Infectious bursal disease virus (IBDV), a member of the Birnaviridae family, is a major avian pathogen responsible for an immunosuppressive disease affecting juvenile chickens. The IBDV genome is formed by two dsRNA segments. The largest one harbors two partially overlapping open reading frames encoding a non-structural polypeptide, known as VP5, and a large polyprotein, respectively. VP5 is non-essential for virus replication. However, it plays a major role in IBDV pathogenesis. VP5 accumulates at the plasma membrane (PM) of IBDV-infected cells. We have analyzed the mechanism underlying the VP5 PM targeting. Updated topological prediction algorithm servers fail to identify a transmembrane domain within the VP5 sequence. However, the VP5 polycationic C-terminal region, harboring three closely spaced patches formed by two or three consecutive basic amino acid residues (lysine or arginine), might account for its PM tropism. We have found that mutations, either C-terminal VP5 deletions or replacement of basic amino acids by alanine residues, that reduce the electropositive charge of the VP5 C-terminus abolish PM targeting. Lipid overlay assays performed with an affinity-purified Flag-tagged VP5 (FVP5) protein version show that this polypeptide binds several phosphoinositides (PIP), exhibiting a clear preference for monophosphate species. Experiments performed with FVP5 mutant proteins lacking the polycationic domain demonstrate that this region is essential for PIP binding. Data gathered with IBDV mutants expressing C-terminal deleted VP5 polypeptides generated by reverse genetics demonstrate that the VP5-PIP binding domain is required both for its PM targeting in infected cells, and for efficient virus dissemination. Data presented here lead us to hypothesize that IBDV might use a non-lytic VP5-dependent cell-to-cell spreading mechanism.
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影响因子：
64.8
作者：
Feng Z;Hensley L;McKnight KL;Hu F;Madden V;Ping L;Jeong SH;Walker C;Lanford RE;Lemon SM
通讯作者：
Lemon SM
影响因子：
5.6
作者：
Luque D;Saugar I;Rejas MT;Carrascosa JL;Rodríguez JF;Castón JR
通讯作者：
Castón JR
影响因子：
1.6
作者：
Hernandez, Martin;Villegas, Pedro;Perez, Ruben
通讯作者：
Perez, Ruben
DOI：
10.1093/protein/15.9.745
发表时间：
2002-09-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Cserzö, M;Eisenhaber, F;Simon, I
通讯作者：
Simon, I
影响因子：
5.6
作者：
Käll, L;Krogh, A;Sonnhammer, ELL
通讯作者：
Sonnhammer, ELL