SARS-CoV-2 and Autophagy

项目概要/摘要 冠状病毒是世界范围内疾病的主要原因，从普通感冒到引起疾病的菌株爆发， 严重的，有时是致命的呼吸窘迫，包括SARS，MERS，以及目前爆发的COVID-1， 19例由SARS-CoV-2引起。在过去的二十年里，人们观察到多种冠状病毒 与自噬途径的成分相互作用，重新排列细胞膜，并产生 RNA复制在这个应用中，我们建议测试SARS-CoV-2与自噬的相互作用， 通路我们将快速评估SARS-CoV-2是否与自噬途径有类似的相互作用， 其他冠状病毒，然后通过确定这些相互作用如何干扰病毒复制， 提供新的治疗靶点。我们将通过三个目标来实施这一战略：第一，我们将 确定SARS-CoV-2复制的早期自噬途径的要求。冠状病毒的数据 研究表明，自噬蛋白LC 3被募集以产生病毒的双膜囊泡， 基因组RNA复制其他数据表明LC 3通过ER相关降解（ERAD）被利用。 制造复制囊泡的机器。我们将确定这两种途径中的哪一种，如果有一种， SARS-CoV-2.在第二个目标中，我们将确定囊泡酸化在SARS-CoV-2复制中的作用。 有强有力的证据表明囊泡的酸化在CoV复制的多个步骤中起作用，我们 将确定囊泡酸化在SARS-CoV-2复制中的作用。在第三个目标中，我们将确定 SARS-CoV-2非结构蛋白在诱导自噬机制中的特定作用。我们有 确定了先前关于冠状病毒中非结构蛋白表达的研究，并将以此指导我们的 SARS-CoV-2蛋白的研究。这些目标补充了RNA病毒生命周期中自噬的现有研究 由杰克逊实验室，同时利用现有的专业知识，弗里曼实验室在研究细胞生物学， 冠状病毒感染。通过完成这些目标，我们将了解SARS-CoV-2是否与 自噬途径以类似或独特的方式与其他冠状病毒相比，并延长了 通过识别可能用于COVID-19治疗的特定宿主-病毒相互作用，在冠状病毒领域进行研究。

PROJECT SUMMARY/ABSTRACT Coronaviruses are major causes of disease worldwide, from the common cold to outbreaks of strains causing severe and sometimes lethal respiratory distress, including SARS, MERS, and the current outbreak of COVID- 19 caused by SARS-CoV-2. It has been observed over the last two decades that multiple coronaviruses interact with components of the autophagy pathway to rearrange cellular membranes and generate sites for RNA replication. In this application, we propose to test the interactions of SARS-CoV-2 with the autophagy pathway. We will rapidly assess whether SARS-CoV-2 has similar interactions with the autophagy pathway as other coronaviruses, then extend the field by identifying how these interactions interfere with viral replication, providing novel therapeutic targets. We will carry this strategy out through three Aims: In the first, we will identify requirements from the early autophagy pathway for SARS-CoV-2 replication. Data from coronavirus studies suggest that the autophagy protein LC3 is recruited to generate double membraned vesicles for virus genomic RNA replication. Other data suggest LC3 is utilized through the ER-associated degradation (ERAD) machinery to generate replication vesicles. We will determine which, if either, of these two pathways is used by SARS-CoV-2. In the second aim, we will determine the role of vesicle acidification in SARS-CoV-2 replication. There is strong evidence that acidification of vesicles plays a role in multiple steps of CoV replication, and we will identify the role of vesicle acidification in SARS-CoV-2 replication. In the third aim, we will identify the specific roles of SARS-CoV-2 non-structural proteins in induction of the autophagic machinery. We have identified previous studies of expression of non-structural proteins in coronavirus and will use this to guide our study of SARS-CoV-2 proteins. These Aims complement existing studies of autophagy in RNA viral life cycles by the Jackson Lab while leveraging the existing expertise of the Frieman Lab in studying cell biology of coronavirus infections. By completing these Aims, we will understand whether SARS-CoV-2 interacts with the autophagy pathway in a similar or unique fashion compared to other coronaviruses, and extend the coronavirus field by identifying specific host-virus interactions that may be targeted for COVID-19 therapies.