Roles of the UL148 glycoprotein in human cytomegalovirus cell tropism

UL148 糖蛋白在人巨细胞病毒细胞趋向性中的作用

基本信息

项目摘要

 DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) is a herpesvirus pathogen that causes life threatening infections in people with compromised immune systems, and congenital defects in newborn children. HCMV depends on its viral envelope glycoproteins to infect human cells. A pair of viral glycoproteins, gH and gL, form a complex, gH/gL, that plays crucial roles in entry of HCMV. As is also the case for Epstein Barr virus (EBV) and human herpesvirus 6 (HHV-6), HCMV virions contain two alternative forms of gH/gL. In HCMV, one of these is a complex with gO, gH/gL/gO, and the other is a complex with UL128, UL130 and UL131, gH/gL/UL128-131. The gH/gL/gO complex suffices for infection of fibroblasts, while gH/gL/UL128-131 is necessary for infection of epithelial and endothelial cells. Viral factors that regulate the relative abundance of alternative gH/gL complexes have not been previously described for any herpesvirus. Intriguingly, we have discovered that HCMV encodes an ER-resident glycoprotein, UL148, which may represent the first example of such a factor. UL148 strongly influences the relative amounts of the two gH/gL complexes on virions, and has large effects on cell tropism. The Specific Aims of this project are to define the influence of UL148 on cell tropism and on maturation of gH/gL complexes, and to identify and characterize protein-protein interactions involving UL148. To carry out these aims, UL148 null mutants of multiple HCMV strains will be characterized, and whether variation in the UL148 gene contributes to the well-documented differences in tropism between HCMV strains will be determined. Studies will also be conducted to determine the effects on tropism and gH/gL when the UL148 is restored to laboratory-adapted HCMV strain AD169, which otherwise lacks the gene. Proteins that co-purify with UL148 during immunoprecipitation from infected cells will be identified by Western blotting and mass spectrometry. Protein interactions involving UL148 and its effects on maturation of gH/gL complexes will be characterized in uninfected cells ectopically expressing the proteins. Chimeric UL148 proteins, in which specific domains of UL148 are swapped with topologically similar proteins, and mutant UL148 proteins, lacking either its ectodomain or cytoplasmic tail, will be used to define the domains necessary for its functions and interactions. Recombinant viruses will be constructed using infectious bacterial artificial chromosome clones of HCMV and sequence manipulation techniques in E. coli. Western blotting, pulse-chase labeling, and enzymatic approaches will be used to visualize the influence of UL148 on gH/gL complexes. Cell tropism will be measured by comparing infectivity on epithelial cells, endothelial cells and fibroblasts. Completion of the project will result in new information that addresses gaps in the current understanding of how HCMV cell tropism is regulated. The resulting findings may have implications for HCMV vaccines, and for understanding the cell tropism of other medically important herpesviruses, such as HHV-6 and EBV, which likewise deploy alternative gH/gL complexes.
 描述(由申请方提供):人巨细胞病毒(HCMV)是一种疱疹病毒病原体,可导致免疫系统受损人群的危及生命的感染和新生儿的先天性缺陷。HCMV依赖于其病毒包膜糖蛋白感染人类细胞。一对病毒糖蛋白gH和gL形成复合物gH/gL,其在HCMV进入中起关键作用。与爱泼斯坦巴尔病毒(EBV)和人疱疹病毒6型(HHV-6)的情况一样,HCMV病毒体含有两种替代形式的gH/gL。在HCMV中,其中一种是与gO的复合物,gH/gL/gO,另一种是与UL 128、UL 130和UL 131的复合物,gH/gL/UL 128 -131。gH/gL/gO复合物足以感染成纤维细胞,而gH/gL/UL 128 -131是感染上皮细胞和内皮细胞所必需的。调节替代性gH/gL复合物相对丰度的病毒因子先前未描述于任何疱疹病毒。有趣的是,我们已经发现HCMV编码ER驻留糖蛋白UL 148,这可能是这种因子的第一个例子。UL 148强烈影响病毒粒子上两种gH/gL复合物的相对量,并且对细胞嗜性具有较大影响。本项目的具体目的是确定UL 148对细胞向性和gH/gL复合物成熟的影响,并鉴定和表征涉及UL 148的蛋白质-蛋白质相互作用。为了实现这些目标,将对多种HCMV毒株的UL 148无效突变体进行表征,并确定UL 148基因的变异是否有助于HCMV毒株之间的嗜性差异。还将进行研究,以确定当将UL 148恢复到实验室适应的HCMV毒株AD 169时对嗜性和gH/gL的影响,否则将缺乏该基因。在免疫沉淀过程中与UL 148共纯化的来自感染细胞的蛋白质将通过蛋白质印迹和质谱法鉴定。涉及UL 148的蛋白质相互作用及其对gH/gL复合物成熟的影响将在异位表达蛋白质的未感染细胞中表征。嵌合UL 148蛋白,其中UL 148的特定结构域与拓扑相似的蛋白质交换,和突变体UL 148蛋白,缺乏其胞外域或胞质尾,将用于定义其功能和相互作用所需的结构域。重组病毒的构建将利用HCMV的感染性细菌人工染色体克隆和E.杆菌Western印迹、脉冲追踪标记和酶促方法将用于可视化UL 148对gH/gL复合物的影响。将通过比较对上皮细胞、内皮细胞和成纤维细胞的感染性来测量细胞嗜性。该项目的完成将产生新的信息,解决目前对HCMV细胞嗜性如何调节的理解中的差距。由此产生的发现可能对HCMV疫苗以及理解其他医学上重要的疱疹病毒(如HHV-6和EBV)的细胞嗜性具有影响,这些病毒同样部署了替代的gH/gL复合物。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

Jeremy Phillip Kamil其他文献

Jeremy Phillip Kamil的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('Jeremy Phillip Kamil', 18)}}的其他基金

Roles of the UL148 glycoprotein in human cytomegalovirus cell tropism
UL148 糖蛋白在人巨细胞病毒细胞趋向性中的作用
  • 批准号:
    9029924
  • 财政年份:
    2015
  • 资助金额:
    $ 36.25万
  • 项目类别:
Roles of the UL148 glycoprotein in human cytomegalovirus infection
UL148糖蛋白在人巨细胞病毒感染中的作用
  • 批准号:
    10735696
  • 财政年份:
    2015
  • 资助金额:
    $ 36.25万
  • 项目类别:
RB POCKET PROTEINS AND E2F TRANSCRIPTION FACTORS IN HCMV REPLICATION
HCMV 复制中的 RB 口袋蛋白和 E2F 转录因子
  • 批准号:
    8359698
  • 财政年份:
    2011
  • 资助金额:
    $ 36.25万
  • 项目类别:
Role of the Human Cytomegalovirus UL97 Proein Kinase in Viral Replication
人巨细胞病毒 UL97 蛋白激酶在病毒复制中的作用
  • 批准号:
    7546053
  • 财政年份:
    2008
  • 资助金额:
    $ 36.25万
  • 项目类别:
Role of the Human Cytomegalovirus UL97 Proein Kinase in Viral Replication
人巨细胞病毒 UL97 蛋白激酶在病毒复制中的作用
  • 批准号:
    7640579
  • 财政年份:
    2008
  • 资助金额:
    $ 36.25万
  • 项目类别:

相似海外基金

How Does Particle Material Properties Insoluble and Partially Soluble Affect Sensory Perception Of Fat based Products
不溶性和部分可溶的颗粒材料特性如何影响脂肪基产品的感官知觉
  • 批准号:
    BB/Z514391/1
  • 财政年份:
    2024
  • 资助金额:
    $ 36.25万
  • 项目类别:
    Training Grant
BRC-BIO: Establishing Astrangia poculata as a study system to understand how multi-partner symbiotic interactions affect pathogen response in cnidarians
BRC-BIO:建立 Astrangia poculata 作为研究系统,以了解多伙伴共生相互作用如何影响刺胞动物的病原体反应
  • 批准号:
    2312555
  • 财政年份:
    2024
  • 资助金额:
    $ 36.25万
  • 项目类别:
    Standard Grant
RII Track-4:NSF: From the Ground Up to the Air Above Coastal Dunes: How Groundwater and Evaporation Affect the Mechanism of Wind Erosion
RII Track-4:NSF:从地面到沿海沙丘上方的空气:地下水和蒸发如何影响风蚀机制
  • 批准号:
    2327346
  • 财政年份:
    2024
  • 资助金额:
    $ 36.25万
  • 项目类别:
    Standard Grant
Graduating in Austerity: Do Welfare Cuts Affect the Career Path of University Students?
紧缩毕业:福利削减会影响大学生的职业道路吗?
  • 批准号:
    ES/Z502595/1
  • 财政年份:
    2024
  • 资助金额:
    $ 36.25万
  • 项目类别:
    Fellowship
感性個人差指標 Affect-X の構築とビスポークAIサービスの基盤確立
建立个人敏感度指数 Affect-X 并为定制人工智能服务奠定基础
  • 批准号:
    23K24936
  • 财政年份:
    2024
  • 资助金额:
    $ 36.25万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Insecure lives and the policy disconnect: How multiple insecurities affect Levelling Up and what joined-up policy can do to help
不安全的生活和政策脱节:多种不安全因素如何影响升级以及联合政策可以提供哪些帮助
  • 批准号:
    ES/Z000149/1
  • 财政年份:
    2024
  • 资助金额:
    $ 36.25万
  • 项目类别:
    Research Grant
How does metal binding affect the function of proteins targeted by a devastating pathogen of cereal crops?
金属结合如何影响谷类作物毁灭性病原体靶向的蛋白质的功能?
  • 批准号:
    2901648
  • 财政年份:
    2024
  • 资助金额:
    $ 36.25万
  • 项目类别:
    Studentship
Investigating how double-negative T cells affect anti-leukemic and GvHD-inducing activities of conventional T cells
研究双阴性 T 细胞如何影响传统 T 细胞的抗白血病和 GvHD 诱导活性
  • 批准号:
    488039
  • 财政年份:
    2023
  • 资助金额:
    $ 36.25万
  • 项目类别:
    Operating Grants
New Tendencies of French Film Theory: Representation, Body, Affect
法国电影理论新动向:再现、身体、情感
  • 批准号:
    23K00129
  • 财政年份:
    2023
  • 资助金额:
    $ 36.25万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
The Protruding Void: Mystical Affect in Samuel Beckett's Prose
突出的虚空:塞缪尔·贝克特散文中的神秘影响
  • 批准号:
    2883985
  • 财政年份:
    2023
  • 资助金额:
    $ 36.25万
  • 项目类别:
    Studentship
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了