Visualization of Influenza Viral RNA Assembly

流感病毒 RNA 组装的可视化

基本信息

批准号：
10681630
负责人：
Seema S. Lakdawala
金额：
$ 38.16万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-11 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10681630
关键词：
Address Binding Proteins Biochemical Biochemistry Biology Biotin Biotinylation Birds Cell Culture System Cell Culture Techniques Cell Nucleus Cell membrane Cell model Cells Cessation of life Color Custom Cytoskeletal Proteins Data Endosomes Epidemic Epithelial Cells Event Family suidae Fluorescent in Situ Hybridization Future Genetic Genetic Materials Genetic Processes Genomic approach Goals Hospitalization Human Imaging Techniques Infection Influenza Influenza A Virus, H1N1 Subtype Influenza A virus Integration Host Factors Intracellular Transport Knowledge Light Location Methods Microscope Microscopy Microtubules Modeling Molecular Virology Movement Nature Nuclear Export Organelles Pathway interactions Pattern Physiological Plasma Play Population Primary Cell Cultures Process Proteins Public Health Publishing RNA RNA Transport Reassortant Viruses Recycling Research Research Personnel Role Route Study models Techniques Viral Viral Genome Viral Proteins Virion Virus Virus Assembly Virus Diseases Virus Replication Visualization Work Zoonoses airway epithelium base cell transformation cell type co-infection imaging approach influenza infection influenzavirus insight live cell imaging new therapeutic target novel nucleocytoplasmic transport pandemic disease pandemic influenza respiratory swine flu therapeutic target tool trafficking viral RNA virology

项目摘要

Influenza A viruses (IAV) pose a major public health threat through both seasonal epidemics and sporadic pandemics. The segmented nature of the viral genome promotes reassortment, a process where the genetic material between viruses is exchanged in a co-infected cell. In nature, reassortment leads to increased viral diversity and emergence of pandemic influenza viruses. For example, the 2009 influenza H1N1 (‘swine flu’) pandemic virus, emerged from reassortment of two circulating swine viruses. Prediction of future pandemic influenza viruses from circulating zoonotic virus populations is difficult because very little is known about the mechanism of reassortment within a single co-infected cell. To accurately define the process of reassortment, we must first understand the dynamics of intracellular viral RNA (vRNA) assembly. Influenza vRNA replicates in the nucleus and is transported to the plasma membrane for packaging, which requires one copy of all eight segments to assemble within a single virion to produce a fully infectious virus. In this proposal, we will build upon our previous data on influenza assembly and define 1) the assembly dynamics in physiologically relevant human and swine cell types, 2) the cellular proteins modulating vRNA transport, and 3) the location of reassortment within a co-infected cell. Our central hypothesis is that vRNA assembly occurs in a cell-type specific manner that correlates with IAV reassortment in different host species. The Specific Aims of this application will use a variety of sophisticated microscopy tools, including live cell imaging with a custom light-sheet microscope, to determine the assembly mechanism in various cell culture models. Aim 1 will utilize multicolor fluorescent in situ hybridization and live cell imaging techniques to explore the dynamics of influenza vRNA assembly in human and swine differentiated airway epithelial cells. Aim 2 will uncover the identity and roles of cellular cytoskeletal proteins and membranous organelles utilized during influenza vRNA assembly using biochemical approaches like proximity-dependent biotinylation. Aim 3 will combine imaging and genomic approaches to characterize the cellular location of vRNA intermingling during co-infection with two heterologous viruses in differentiated airway epithelial cells. The proposed work will address many outstanding questions in influenza biology regarding reassortment that have remained unanswered due to a lack of tools to track vRNA movement in live cells during a productive infection. In addition, these studies will identify novel host factors involved in vRNA packaging that can be pursued as potential therapeutic targets.

流感的病毒（IAV）RAL基因组促进了REASTREMT，在自然界中，病毒之间的遗传物质是在自然界中交换的2009年流感H1N1和病毒，对循环IC病毒种群的未来大流行性流感病毒的预测很困难，因为很少有Ikout在单个共同感染的细胞中的重新分配机制，我们必须首先了解细胞内病毒RNA（VRNA）组件的动态学。。我们将基于先前的数据Onza组装并定义1）生理性相关的人和猪细胞类型中的组装动力学，2）细胞蛋白vrna转化O感染的细胞。与不同宿主物种中的IAV重新出现相关的特定方式杂交和活细胞成像与人类和猪的动力学在人和猪的上皮细胞中使用eNza vrna组装。由于在vrna包装中缺乏otrack vrna运动，因此可以将其作为潜在的治疗靶标的释放。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

mSphere of Influence: Redefining an Influenza Virus-How Different Are Influenza Viruses and Extracellular Vesicles?

影响范围：重新定义流感病毒——流感病毒和细胞外囊泡有何不同？

DOI：
10.1128/msphere.00563-19
发表时间：
2019
期刊：
mSphere
影响因子：
4.8
作者：
Lakdawala,SeemaS
通讯作者：
Lakdawala,SeemaS

Learning the sequence of influenza A genome assembly during viral replication using point process models and fluorescence in situ hybridization.

使用点过程模型和荧光原位杂交了解病毒复制过程中甲型流感基因组组装的序列。

DOI：
10.1371/journal.pcbi.1006199
发表时间：
2019
期刊：
PLoS computational biology
影响因子：
4.3
作者：
Majarian,TimothyD;Murphy,RobertF;Lakdawala,SeemaS
通讯作者：
Lakdawala,SeemaS

Cell-Culture Adaptation of H3N2 Influenza Virus Impacts Acid Stability and Reduces Airborne Transmission in Ferret Model.

DOI：
10.3390/v13050719
发表时间：
2021-04-21
期刊：
Viruses
影响因子：
0
作者：
Le Sage V;Kormuth KA;Nturibi E;Lee JM;Frizzell SA;Myerburg MM;Bloom JD;Lakdawala SS
通讯作者：
Lakdawala SS

Teaching an Old Virus New Tricks: A Review on New Approaches to Study Age-Old Questions in Influenza Biology.