Membrane Remodeling in Viral Infection and Viral Assembly

病毒感染和病毒组装中的膜重塑

• 批准号: 10920195
• 负责人: JOSHUA ZIMMERBERG
• 金额: $ 139.54万
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10920195
• 关键词: Agreement; Amino Acids; Antigens; Cell membrane; Cells; Chemistry; Cytoplasm; Data; Electron Microscopy; Experimental Models; Glycoproteins; Hemagglutinin; Hybrids; Hydration status; Hydrophobic Surfaces; Hydrophobicity; In Vitro; Infection; Influenza; Lateral; Lead; Lipid Bilayers; Lipids; Mediating; Membrane; Membrane Fusion; Membrane Lipids; Modeling; Molecular Conformation; Morbidity - disease rate; N-terminal; Paper; Pathway interactions; Peptides; Phospholipids; Probability; Proteins; Rabies; Reporting; Rest; Role; Rubella; Rupture; Side; Structure; Surface; System; Tail; Tertiary Protein Structure; Therapeutic; Thick; Thinness; Transmembrane Domain; Transmembrane Transport; Variant; Viral; Viral Fusion Proteins; Viral Genome; Virus; Virus Assembly; Virus Diseases; Visualization; Water; amphiphilicity; dimer; in vivo; influenza virus strain; influenzavirus; membrane model; molecular dynamics; mortality; receptor; restraint; simulation

In the first paper reporting this discovery, a study of the membrane mechanisms by which the influenza virus can disrupt a target membrane, we establish that FPs underly this disruption: in target membranes, a reversible pore forms upon addition of FP in the absence of virus or even the rest of HA. In MD simulations crafted to understand the chemistry by which FPs act, a third kind of rosette emerged: the aggregation of FP via their lateral side chains (not their hydrophobic surfaces) into FP microdomains that displace lipids in the cis leaflet. This aggregated structure locally thinned the bilayer and significantly increased the probability of water entry. A new model is proposed to explain our data based on a tilting of FPs towards each other to further thin the remaining lipids immediately under even an FP dimer. For larger aggregates, this more hydrated, thinner membrane structure replaces the lipid bilayer in a small domain wherein a lipidic pore can form. A second paper on the membrane mechanisms by which the influenza virus can disrupt a target membrane. The fusion peptide (FP) domain is necessary for the fusogenic activity of spike proteins in a variety of enveloped viruses, allowing the virus to infect the host cell, and is the only part of the protein that interacts directly with the target membrane lipid tails during fusion. There are consistent findings of poration by this domain in experimental model membrane systems, and, in certain conditions, the isolated FPs can generate pores. Here, we use molecular dynamics simulations to investigate the specifics of how these FP-induced pores form in membranes with different compositions of lysolipid and POPC. The simulations show that pores form spontaneously at high lysolipid concentrations via hybrid intermediates, where FP aggregates in the cis leaflet tilt to form a funnel-like structure that spans the leaflet and locally reduces the hydrophobic thickness that must be traversed by water to form a pore. By restraining a single FP within an FP aggregate to this tilted conformation, pores can be formed in lower-lysolipid-content membranes, including pure POPC, on the 100-ns timescale, much more rapidly than in unbiased simulations in bilayers with the same composition. The pore formation pathway is similar to the spontaneous formation in high lysolipid concentrations. Depending on the membrane composition, the pores can be metastable (as seen in POPC) or lead to membrane rupture.

在报告这一发现的第一篇论文中，研究了流感病毒可以破坏靶膜的膜机制，我们确定了FP是这种破坏的基础：在靶膜中，在没有病毒或甚至没有HA的情况下加入FP后形成可逆的孔。在MD模拟中，为了理解FP的化学作用，出现了第三种玫瑰花结：FP通过其侧链（而不是其疏水表面）聚集成FP微结构域，取代顺式小叶中的脂质。这种聚集结构局部地使双层变薄，并显著增加了水进入的可能性。提出了一种新的模型来解释我们的数据的基础上倾斜的FP彼此进一步薄的剩余的脂质下立即甚至FP二聚体。对于较大的聚集体，这种水合程度更高、更薄的膜结构取代了小结构域中的脂质双层，在该结构域中可以形成疏水孔。 第二篇关于流感病毒破坏靶膜的膜机制的论文。融合肽（FP）结构域是多种包膜病毒中刺突蛋白的融合活性所必需的，允许病毒感染宿主细胞，并且是在融合期间与靶膜脂质尾直接相互作用的蛋白质的唯一部分。在实验模型膜系统中，该结构域的穿孔作用有一致的发现，并且在某些条件下，分离的FP可以产生孔。在这里，我们使用分子动力学模拟来研究这些FP诱导的孔如何在具有不同组成的溶血脂质和POPC的膜中形成的细节。模拟显示，孔通过混合中间体在高溶解脂质浓度下自发形成，其中顺式小叶中的FP聚集体倾斜以形成漏斗状结构，该结构跨越小叶并局部降低必须被水穿过以形成孔的疏水厚度。通过将FP聚集体内的单个FP限制为这种倾斜构象，可以在100 ns时间尺度上在低溶血脂质含量的膜（包括纯POPC）中形成孔，比具有相同组成的双层中的无偏模拟快得多。孔的形成途径类似于高溶脂条件下的自发形成 浓度的根据膜的组成，孔可以是亚稳态的（如POPC中所见）或导致膜破裂。

项目成果

Cytoplasmic free Ca2+ is essential for multiple steps in malaria parasite egress from infected erythrocytes.

• DOI: 10.1186/1475-2875-12-41
• 发表时间: 2013-01-30
• 期刊: Malaria journal
• 影响因子: 3
• 作者: Glushakova S;Lizunov V;Blank PS;Melikov K;Humphrey G;Zimmerberg J
• 通讯作者: Zimmerberg J

Cholesterol promotes hemifusion and pore widening in membrane fusion induced by influenza hemagglutinin.

• DOI: 10.1085/jgp.200709932
• 发表时间: 2008-05
• 期刊: JOURNAL OF GENERAL PHYSIOLOGY
• 影响因子: 3.8
• 作者: Biswas, Subrata;Yin, Shu-Rong;Blank, Paul S.;Zimmerberg, Joshua
• 通讯作者: Zimmerberg, Joshua

Poxvirus membrane biogenesis: rupture not disruption.

• DOI: 10.1111/cmi.12072
• 发表时间: 2013-02
• 期刊: Cellular microbiology
• 影响因子: 3.4
• 作者: Krijnse Locker J;Chlanda P;Sachsenheimer T;Brügger B
• 通讯作者: Brügger B

ER biogenesis: self-assembly of tubular topology by protein hairpins.

ER 生物发生：通过蛋白质发夹进行管状拓扑的自组装。

• DOI: 10.1016/j.cub.2008.04.031
• 发表时间: 2008
• 期刊: Current biology : CB
• 作者: Shnyrova,Anna;Frolov,VadimA;Zimmerberg,Joshua
• 通讯作者: Zimmerberg,Joshua

Nanoscale 3D cellular imaging by axial scanning transmission electron tomography.

纳米级3D细胞成像通过轴向扫描透射电子断层扫描。

• DOI: 10.1038/nmeth.1367
• 发表时间: 2009-10
• 期刊: NATURE METHODS
• 影响因子: 48
• 作者: Hohmann-Marriott, Martin F.;Sousa, Alioscka A.;Azari, Afrouz A.;Glushakova, Svetlana;Zhang, Guofeng;Zimmerberg, Joshua;Leapman, Richard D.
• 通讯作者: Leapman, Richard D.