CAREER: Structural Interplay Between Chromatin Remodeling and Lentiviral Integration

职业：染色质重塑和慢病毒整合之间的结构相互作用

• 批准号: 2048095
• 负责人: Dmitry Lyumkis
• 金额: $ 179.21万
• 依托单位: The Salk Institute for Biological Studies
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2048095&HistoricalAwards=false
• 关键词: CAREER; Structural; Interplay; Between; Chromatin

This research will uncover how some viruses like HIV hijack host protein machinery and permanently alter the host genome to sustain infection. The work will produce atomic structures that reveal the interactions between viral and host proteins that mediate the integration of viral genetic information into the host genome. The resulting structures and biochemical data will provide a mechanistic framework for understanding how these interactions lead to productive infection and will reveal insights into how this process may be blocked, and lead to novel anti-virals, or can be exploited for gene therapy. These research themes will be taught to audiences at different stages of their scientific development. The integrated research and education plan includes the advanced scientific training of both undergraduate and graduate students, the exposure of students to complex scientific programs within a world-renowned academic research institution, and the introduction of both structural biology and virology to under-privileged high-school students to inspire their scientific development. All lentiviruses, including HIV, are characterized by an important hallmark that distinguishes them from other viruses — their ability to integrate a complementary DNA copy of their RNA genome into target host chromatin through a catalytic process called integration. Integration establishes a permanent and irreversible infection in the target cell and contributes to viral pathogenicity. Although we understand the basic mechanisms underlying lentiviral integration, we know comparatively little about the role that host factors play in this process. Specifically, there is the prominent but wholly underappreciated phenomenon of chromatin remodeling that accompanies successful integration. Disruption of host chromatin remodelers leads to distinct phenotypes affecting lentiviral infection, but how the remodelers are assembled, engage chromatin, and ultimately interplay with lentiviral components to affect integration remain unclear. Using cryogenic electron microscopy and biochemical methods, this research will investigate: (1) how ATP-independent chromatin remodelers reorganize chromatin and exploit this reorganized state to engage viral machinery prior to catalytic integration; and (2) how ATP-dependent chromatin remodelers engage and reorganize integrated viral DNA after catalytic integration to establish a transcriptionally competent provirus. This work will provide a mechanistic molecular framework that elucidates how host chromatin remodelers interact with viral machinery to facilitate integration and establish a foundation for future work that broadly examines the structure, function, and cross-talk between the molecular machines responsible for chromatin remodeling and lentiviral integration.This work was jointly funded by the Molecular Biophysics and Genetic Mechanisms clusters.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这项研究将揭示一些病毒如艾滋病毒如何劫持宿主蛋白质机制，并永久改变宿主基因组以维持感染。这项工作将产生原子结构，揭示病毒和宿主蛋白质之间的相互作用，介导病毒遗传信息整合到宿主基因组中。由此产生的结构和生物化学数据将提供一个机制框架，以了解这些相互作用如何导致生产性感染，并将揭示如何阻止这一过程的见解，并导致新的抗病毒药物，或可用于基因治疗。这些研究主题将在其科学发展的不同阶段教授给观众。综合研究和教育计划包括本科生和研究生的高级科学培训，学生在世界知名的学术研究机构内接触复杂的科学课程，并向贫困高中生介绍结构生物学和病毒学，以激发他们的科学发展。 所有的慢病毒，包括HIV，都有一个重要的特征，将它们与其他病毒区分开来-它们能够通过称为整合的催化过程将其RNA基因组的互补DNA拷贝整合到靶宿主染色质中。整合在靶细胞中建立永久和不可逆的感染，并有助于病毒致病性。虽然我们了解慢病毒整合的基本机制，但我们对宿主因素在这一过程中所起的作用知之甚少。具体来说，有一个突出的，但完全低估的现象，染色质重塑，伴随着成功的整合。宿主染色质重塑的破坏导致影响慢病毒感染的不同表型，但重塑如何组装，接合染色质，并最终与慢病毒组分相互作用以影响整合仍不清楚。使用低温电子显微镜和生物化学方法，本研究将调查：（1）ATP-非依赖性染色质重塑如何重组染色质，并利用这种重组状态，从事催化整合前的病毒机制;（2）ATP-依赖性染色质重塑如何参与和重组整合后的病毒DNA催化整合，以建立一个转录能力的前病毒。这项工作将提供一个机制的分子框架，阐明宿主染色质重塑如何与病毒机制相互作用，以促进整合，并为未来的工作奠定基础，广泛研究结构，功能，和交叉在负责染色质重塑和慢病毒整合的分子机器之间进行讨论。这项工作由分子生物物理学和遗传机制集群共同资助。该奖项反映了NSF的基金会的使命是履行其法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Multivalent interactions essential for lentiviral integrase function.

• DOI: 10.1038/s41467-022-29928-8
• 发表时间: 2022-05-03
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Ballandras-Colas, Allison;Chivukula, Vidya;Gruszka, Dominika T.;Shan, Zelin;Singh, Parmit K.;Pye, Valerie E.;McLean, Rebecca K.;Bedwell, Gregory J.;Li, Wen;Nans, Andrea;Cook, Nicola J.;Fadel, Hind J.;Poeschla, Eric M.;Griffiths, David J.;Vargas, Javier;Taylor, Ian A.;Lyumkis, Dmitry;Yardimci, Hasan;Engelman, Alan N.;Cherepanov, Peter
• 通讯作者: Cherepanov, Peter

Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon.

• DOI: 10.3791/64136
• 发表时间: 2022-07-01
• 期刊: Journal of visualized experiments : JoVE
• 作者: Aiyer S;Strutzenberg TS;Bowman ME;Noel JP;Lyumkis D
• 通讯作者: Lyumkis D