Genome-wide mutagenesis reveals that ORF7 is a novel VZV skin-tropic factor.
Genome-wide mutagenesis reveals that ORF7 is a novel VZV skin-tropic factor.
复制标题
DOI：
10.1371/journal.ppat.1000971
复制
发表时间：
2010-07-01
期刊：
影响因子：
6.7
通讯作者：
Zhu H
中科院分区：
文献类型：
--
作者：
Zhang Z;Selariu A;Warden C;Huang G;Huang Y;Zaccheus O;Cheng T;Xia N;Zhu H
水痘带状疱疹病毒（VZV）是一种普遍存在的人类α疱疹病毒，是水痘和带状疱疹的病原体。尽管减毒VZV疫苗（v-Oka）已在美国广泛用于儿童，但水痘疫情仍时有发生，带状疱疹疫苗仅将带状疱疹风险降低50%。因此，VZV仍然是一个重要的公共卫生问题。由于VZV在培养细胞中的高度细胞相关性、产生重组病毒的困难以及VZV对人类细胞和组织的几乎排他性，对VZV复制和发病机制的了解仍然有限。为了绕过这些障碍，我们将整个VZV（p-Oka）基因组克隆到包括双报告系统（GFP和荧光素酶报告基因）的细菌人工染色体中。我们采用PCR诱变和同源重组系统在E。大肠杆菌，分别删除基因组的70个独特的ORF。在MeWo细胞和培养的人胎儿皮肤器官样品中系统地检查所获得的病毒突变体的集合。我们使用我们的全基因组删除库提供新的功能注释的51%的VZV蛋白质组。我们发现70个VZV ORF中有44个是病毒复制所必需的。在26个非必需ORF缺失突变体中，8个在MeWo中具有可辨别的生长缺陷。有趣的是，发现四个ORF是皮肤器官培养物中病毒复制所需的，但不是在MeWo细胞中，这表明它们作为皮肤嗜性因子的潜在作用。其中一个基因（ORF 7）从未被描述为皮肤嗜性因子。VZV基因组的全球分析进一步深入了解了这种病毒的复制和发病机制，这可以改善水痘和带状疱疹的预防和治疗。水痘带状疱疹病毒（VZV）是水痘和带状疱疹的病原体。目前的水痘疫苗的长期有效性尚未确定，目前的带状疱疹疫苗未能为相当多的人提供保护性免疫。带状疱疹还可能导致带状疱疹后神经痛（PHN），这是一种与顽固性疼痛相关的衰弱性疾病，可能会持续一生。因此，VZV仍然是一个重要的公共卫生问题。我们使用我们的全基因组删除库的增长率分析，以确定所有已知的VZV基因的必要性，包括新的注释为51%的VZV蛋白质组。我们还发现了一个新的皮肤嗜性因子编码的ORF 7。总的来说，我们对VZV复制和发病机制所必需的基因的鉴定将作为VZV的多个深入遗传研究的基础，这可以改善水痘和带状疱疹的预防和治疗。例如，必需基因可能是有吸引力的药物靶点，而其缺失导致实质性生长缺陷的基因可能是新型减毒活疫苗的潜在候选者。
The Varicella Zoster Virus (VZV) is a ubiquitous human alpha-herpesvirus that is the causative agent of chicken pox and shingles. Although an attenuated VZV vaccine (v-Oka) has been widely used in children in the United States, chicken pox outbreaks are still seen, and the shingles vaccine only reduces the risk of shingles by 50%. Therefore, VZV still remains an important public health concern. Knowledge of VZV replication and pathogenesis remains limited due to its highly cell-associated nature in cultured cells, the difficulty of generating recombinant viruses, and VZV's almost exclusive tropism for human cells and tissues. In order to circumvent these hurdles, we cloned the entire VZV (p-Oka) genome into a bacterial artificial chromosome that included a dual-reporter system (GFP and luciferase reporter genes). We used PCR-based mutagenesis and the homologous recombination system in the E. coli to individually delete each of the genome's 70 unique ORFs. The collection of viral mutants obtained was systematically examined both in MeWo cells and in cultured human fetal skin organ samples. We use our genome-wide deletion library to provide novel functional annotations to 51% of the VZV proteome. We found 44 out of 70 VZV ORFs to be essential for viral replication. Among the 26 non-essential ORF deletion mutants, eight have discernable growth defects in MeWo. Interestingly, four ORFs were found to be required for viral replication in skin organ cultures, but not in MeWo cells, suggesting their potential roles as skin tropism factors. One of the genes (ORF7) has never been described as a skin tropic factor. The global profiling of the VZV genome gives further insights into the replication and pathogenesis of this virus, which can lead to improved prevention and therapy of chicken pox and shingles. The Varicella Zoster Virus (VZV) is the causative agent of chicken pox and shingles. The long-term efficacy of the current chickenpox vaccine is yet to be determined, and the current shingles vaccine fails to provide protective immunity for a substantial number of individuals. Shingles can also lead to post-herpetic neuralgia (PHN), a debilitating condition associated with an intractable pain that can linger for life. Therefore, VZV remains an important public health concern. We use growth-rate analysis of our genome-wide deletion library to determine the essentiality of all known VZV genes, including novel annotations for 51% of the VZV proteome. We also discovered a novel skin-tropic factor encoded by ORF7. Overall, our identification of genes essential for VZV replication and pathogenesis will serve as the basis for multiple in-depth genetic studies of VZV, which can lead to improved prevention and therapy of chicken pox and shingles. For example, essential genes may be appealing drug targets and genes whose deletion causes a substantial growth defect may be prospective candidates for novel live attenuated vaccines.
登录
查看更多内容
影响因子：
12.3
作者：
Gentleman RC;Carey VJ;Bates DM;Bolstad B;Dettling M;Dudoit S;Ellis B;Gautier L;Ge Y;Gentry J;Hornik K;Hothorn T;Huber W;Iacus S;Irizarry R;Leisch F;Li C;Maechler M;Rossini AJ;Sawitzki G;Smith C;Smyth G;Tierney L;Yang JY;Zhang J
通讯作者：
Zhang J
影响因子：
5.4
作者：
COHEN, JI;SEIDEL, K
通讯作者：
SEIDEL, K
影响因子：
5.4
作者：
Che, Xibing;Berarducci, Barbara;Arvin, Ann M.
通讯作者：
Arvin, Ann M.
DOI：
10.3923/pjbs.2007.2140.2145
发表时间：
2007-07-01
期刊：
Pakistan journal of biological sciences : PJBS
影响因子：
--
作者：
Gharavi, Sara;Sadeghizadeh, Majid;Sabahi, Farzaneh
通讯作者：
Sabahi, Farzaneh
影响因子：
5.4
作者：
Erazo, Angela;Yee, Michael B.;Kinchington, Paul R.
通讯作者：
Kinchington, Paul R.