Dissection of Subcellular Responses and Interactions in Plants

所有生物体都会对环境信号做出反应，并采用各种亚细胞策略来最大限度地提高生存机会。植物，尽管他们的久坐不动的性质已经特别成功地对抗周围的无数压力。因此，植物细胞一直是我实验室的首选研究材料，我们努力了解它们维持细胞稳态的微妙策略。我的长期目标是了解植物细胞的反应性，这项研究是通过我在2005年的第一次发现资助开始的，并无缝地延伸到第二个资助期，从发现加速补充中受益匪浅。在过去的资助期间，我的基于实时成像的方法已经导致了各种各样的基本工具，使用传统的和光转换荧光蛋白。该实验室还开发了几种开创性的技术，用于精确靶向和跟踪亚细胞成分和隔室。最重要的是，牢记我们的长期目标，我们发现并初步表征了涉及来自不同细胞器的快速膜延伸的新亚细胞现象（Sinclair et al.，2009; Mathur等人，2013; Schattat等人，2012）。我们报告的瞬时现象似乎是所有真核生物都共有的，并且经常涉及使应激细胞迅速恢复平衡，以及使其更好地准备应对复发性应激。这个建议建立在我过去10年的基础工作之上，旨在揭示迄今为止确定的基本亚细胞现象的更多细节。我为现时的建议所界定的四个主要和相辅相成的研究范畴是：了解细胞器延伸形成的机制。（二）、了解细胞器延伸在细胞器间运输中的作用。（三）、了解内质网（ER）作为细胞器形状和相互作用的物理调节器的作用。（四）、了解内膜-细胞骨架相互作用的基本原理。

All living organisms respond to environmental cues and employ a variety of sub-cellular strategies for maximizing their chances of survival. Plants, despite their sedentary nature have been especially successful in combating the myriad stresses surrounding them. Plant cells, therefore, have been the research material of choice for my lab where we strive to understand their subtle stratagems for maintaining cellular homeostasis. Research into my long-term goal of understanding the responsiveness of the plant cell was initiated through my first Discovery grant in 2005 and has extended seamlessly into the second grant period with considerable benefit from a Discovery accelerated supplement. During the past grant periods my live-imaging based approach has resulted in a wide variety of basic tools using conventional and photo-convertible fluorescent proteins. The lab has also developed several pioneering techniques for precise targeting and tracking of sub-cellular components and compartments. Most important, keeping our long-term goal in mind, is our discovery and initial characterization of novel sub-cellular phenomenon involving rapid membrane extensions from different organelles (Sinclair et al., 2009; Mathur et al., 2013; Schattat et al. 2012). The transient phenomena reported by us appear to be common to all eukaryotes and are very often involved in bringing a stressed cell back to equilibrium quickly as well as allowing it to be better prepared for dealing with recurrent stress. This proposal builds up on my foundational work of the last 10 years and is aimed at uncovering more details about the basic subcellular phenomena identified so far. The four key and complementary areas of exploration defined by me for investigation under the present proposal are: (1). Understanding the mechanisms involved in the formation of organelle extensions. (2). Understanding the role of organelle extensions in inter-organelle trafficking. (3). Understanding the role of the endoplasmic reticulum (ER) as a physical regulator of organelle shape and interactivity.(4). Understanding the fundamentals of endomembrane-cytoskeletal interactions.