Experimental Model Systems for Intracellular Compartmentalization: Dynamic Formation/Disassembly of Model Organelles in Artificial Cells

细胞内区室化的实验模型系统：人工细胞中模型细胞器的动态形成/分解

基本信息

批准号：
1244180
负责人：
Christine Keating
金额：
$ 81.25万
依托单位：
Pennsylvania State Univ University Park
依托单位国家：
美国
项目类别：
Continuing Grant
财政年份：
2013
资助国家：
美国
起止时间：
2013-03-01 至 2018-02-28
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1244180&HistoricalAwards=false
关键词：
Experimental Model Systems Intracellular Compartmentalization

项目摘要

Technical Description: This proposal is based on the idea that it may be possible to realize key structural and functional aspects of subcellular organization due to the physicochemical phenomena that arise in solutions that are crowded with macromolecules "like the cytoplasm and nucleoplasm are" even in the absence of specific biomolecular interactions. Of particular interest is aqueous phase separation, which commonly occurs in polymer solutions and offers a means of compartmentalization. This work will evaluate two hypotheses: (1) that phase separation in the crowded intracellular environment could be largely responsible for the existence and properties of non-membranous organelles, and (2) that these structures could in turn serve as templates for membrane assembly. Experimental models of subcellular cytoplasmic and nuclear compartments will be developed based on crowded solutions of RNA, polyamines, proteins, and neutral polymers. Protein phosphorylation will be used to drive formation and disassembly of microcompartments that will serve as model organelles. Three research objectives are proposed: (1) Dynamic assembly/disassembly of model non-membranous organelles based on aqueous phase compartments that form in response to protein phosphorylation state. (2) Templated membrane formation around these model organelles. (3) Primitive model for mitosis/cell cycle in cell-sized lipid vesicles that contain the crowded solution and phase compartment-based model organelles. Nontechnical Description: Intracellular organization is a hallmark of living cells, with both membrane-bounded (e.g., nucleus) and non-membranous organelles (e.g., nucleolus, P-granules) performing key cellular functions. The central questions driving these investigations are: What role do relatively nonspecific chemical and physical effects play in subcellular organization and the associated functions of biological cells, and how does complexity arise from a small number of simple molecular components? The investigators hypothesize that, despite the deliberate simplicity of the proposed model cells, they will be able to mimic complex biological processes and specific biochemical interactions, such as the reversible formation and dissolution of RNA and protein-rich compartments, the formation of interior membranes around pre-existing protein-rich compartments, and finally to model cell division in a very primitive mimic of the mitotic cell. Broader Impacts. Two graduate students will be trained at the interface of molecular and cell biology, chemistry, biophysics and materials science. Undergraduate students will be recruited to work on this project during the academic year for course credit and in the summer through the various on-site REU programs, with a target of one student during the academic year and one or two students during the summer. Support for one K-12 teacher each summer is included in the budget. The investigators will team with the existing Research Experiences for Teachers program in the PSU MRSEC. Teachers will perform experiments and develop curriculum materials to bring back to their own classrooms the following school year. Real-world examples go beyond the intracellular organelles that motivate the intellectual merit of this proposal to also include, e.g., food science, drug delivery, and environmental remediation.This award is being funded by the Systems and Synthetic Biology Cluster in MCB/BIO and co-funded by the Chemistry of Life Processes Program in CHE/MPS.

技术描述：该提案是基于这样的想法：由于在挤满大分子的溶液中出现的物理化学现象而产生的物理化学现象可能会实现亚细胞组织的关键结构和功能方面，例如细胞质和核质量，即使缺乏特定的生物分子相互作用。特别感兴趣的是水相分离，通常发生在聚合物溶液中，并提供一种分隔的方法。这项工作将评估两个假设：（1）拥挤的细胞内环境中的相位分离可能在很大程度上负责非膜细胞器的存在和特性，以及（2）这些结构又可以用作膜组装的模板。基于RNA，多胺，蛋白质和中性聚合物的拥挤溶液，将开发亚细胞胞质和核区室的实验模型。蛋白质磷酸化将用于驱动将用作模型细胞器的微型室的形成和拆卸。提出了三个研究目标：（1）基于响应蛋白质磷酸化状态的水相隔室的非膜器隔室的动态组装/拆卸。（2）这些模型细胞器周围的膜形成。（3）包含拥挤溶液和基于相隔室模型细胞器的细胞大小脂质囊泡中有丝分裂/细胞周期的原始模型。非技术描述：细胞内组织是活细胞的标志，具有膜结合的（例如核）和非膜细胞器（例如核仁，核，p颗粒），可执行关键细胞功能。推动这些研究的中心问题是：相对非特异性的化学和物理效应在亚细胞组织以及生物细胞的相关功能中起什么作用？研究者假设，尽管提出的模型细胞有意简单，但它们将能够模仿复杂的生物学过程和特定的生化相互作用，例如富含RNA和富含蛋白质的隔室的可逆形成和溶解，并在富含蛋白质的细胞围绕良好的细胞和模型moim MoIM MIMIM MORIM MORIM MORIIM MORIIM MORIIM MORIM MORIM MOLIM MOLIM MOLIIM MOLIM MOLIM MOLIM MOLIM MOLIM MORIIM MOLIM MORIM MOLIM MOLIIM MONIIM MOLIIM MOLIIM MONIIM MONIM MONIIM MONIM IIM IIM IIM IIM IIM IIM IIM IIM组成。更广泛的影响。两名研究生将接受分子和细胞生物学，化学，生物物理学和材料科学的界面培训。本科生将在学年中招募该项目，以获取课程学分，并在夏天通过各种现场REU课程，在学年中，一个学生的目标是一个学生，在夏季的一两个学生。预算中包括对每年夏天的一名K-12老师的支持。调查人员将与PSU MRSEC中的教师计划的现有研究经验合作。老师将在下一个学年进行实验并开发课程材料，以将其带回自己的教室。现实世界中的例子超出了细胞内细胞器，激发了该提案的智力优点，例如，例如食品科学，药物提供和环境修复。该奖项由MCB/Bio中的系统和合成生物学集群资助，并由CHE/MPS的Life Processions in Che/MPS中的Chemistry Progessions授予。