Elucidating the molecular mechanisms of regulation of cellular stress sensors

对压力的反应是生物体生理和生物学的一个组成部分。在生存的进化斗争中，适应压力环境是一个决定性的优势。内质网（ER）是一个位于细胞中心的细胞器，负责许多家务功能，包括蛋白质和脂质的合成、折叠、细胞内钙的储存和释放。许多内在或外在因素导致内质网功能的破坏最终导致内质网应激，内质网产生应对反应[如未折叠蛋白反应（UPR）]以减轻或消除应激。UPR通路感知内质网干扰，涉及旨在重建蛋白质合成机制的不同组分，包括翻译衰减、编码伴侣和内质网相关降解组分的基因的转录激活，以及凋亡和自噬途径的激活。有三种内质网跨膜应力传感器构成内质网应激的UPR反应：PERK、ATF6和IRE1，它们是具有腔内应激传感结构域的内质网蛋白。我们的研究项目集中于识别参与细胞应激传感器调节的分子机制。本研究的长期目标是产生有关应力传感器分子机制的基础知识，并为参与该项目的HQP提供独特的培训机会。内窥镜环境对UPR的光腔传感器域的调节仍然知之甚少。该研究项目于2014年启动，旨在解决这一知识鸿沟。我们发现内质网驻留蛋白（氧化还原酶，环氧化酶和钙螯合酶）结合到IRE的内质网光应激感应域，影响其细胞信号传导。然而，控制这些相互作用的分子机制尚不清楚。此外，其他内质网应力传感器的调节仍有待研究。为了解决这些问题，我们的研究计划有三个主题，重点是在体外重构机制事件，使用重组IRE1， PERK和ATF6管腔结构域，在相关ER驻留蛋白存在下；并评估内源性环境是否参与了这一过程。主题1。确定ER光应力传感器的性质研究PERK和ATF6 ER应力传感器的分子调控确定内质网应激传感器调控的生物学重要性本研究项目将对内质网应激传感器生物学、分子相互作用在应激传感中的重要性产生新的见解，并推进我们对内质网应激应对反应的一般理解。我们还为HQP提供了良好的培训环境，以满足工业界和学术界对技术和技能的需求。我们成功的长期指标将是学员达到学术研究人员、行业领袖或政府决策者等有影响力的高级职位的百分比。

Responses to stress are an integral part of organism's physiology and biology. Adaptation to stressful conditions is a decisive advantage in the evolutionary struggle for existence. The endoplasmic reticulum (ER) is a centrally located intracellular organelle responsible for many housekeeping functions including the synthesis, folding of proteins and lipids, the storage and release of intracellular calcium. Disruption of ER function caused by many intrinsic or extrinsic factors culminates in ER stress, and the ER mounts a coping response [e.g., unfolded protein response (UPR)] to mitigate or eliminate the stress. The UPR pathways sense ER disturbance and involves distinct components designed to re-establish the protein synthetic machinery, including translational attenuation, transcriptional activation of genes encoding chaperones and components of the ER-associated degradation, and activation of apoptotic and autophagy pathways. There are three ER transmembrane stress sensors that comprise the UPR reaction to ER stress: PERK, ATF6, and IRE1, integral ER proteins with luminal stress sensing domains. Our research program is focused on identifying the molecular mechanisms involved in the regulation of cellular stress sensors. Long-term objectives of this research are to generate fundamental knowledge regarding molecular mechanisms of stress sensors and to provide unique training opportunity for HQP engaged in the program. Regulation of the luminal sensor domain of UPR by the ER luminal environment remains poorly understood. This research program was initiated in 2014 to address this knowledge gap. We discovered that ER resident proteins (oxidoreductase, cyclooxygenase and calsequestrin) bind to ER luminal stress sensing domain of IRE to affect its cellular signaling. However, the molecular mechanisms governing these interactions are not known. Furthermore, regulation of other ER stress sensors remains to be examined. To address these our research program has three themes focused on reconstituting the mechanistic events in vitro, using recombinant IRE1, PERK and ATF6 luminal domain, in the presence of relevant ER resident proteins; and assessing whether ER luminal environment is involved in this process. Theme 1. Determine the nature of ER luminal stress sensors Theme 2. Examine molecular regulation of PERK and ATF6 ER stress sensors Theme 3. Determine biological importance of ER stress sensors regulation This research program will yield new insights on ER stress sensors biology, the importance of molecular interaction in stress sensing, as well as advance our general understanding of ER stress coping responses. We also provide outstanding environment for training of HQP in technologies and skills desired by industry and academic job sectors. Longer-term indicators of our success will be percentages of trainees reaching senior, influential positions as academic researchers, industry leaders or government policy makers.