Bifunctional Control of Yop Translocation by YopK

描述（由申请人提供）：III型分泌是一种通过分子注射器样结构将有毒蛋白直接递送到宿主细胞中的方法，许多革兰氏阴性病原体都采用这种方法。III型分泌系统导致了人类各种各样的疾病，包括黑死病、食源性疾病和囊性纤维化。仅食源性疾病每年就有数百万例，这对我们的卫生保健系统产生了惊人的影响。再加上新出现的抗生素耐药性日益致命的可能性，迫切需要新的抗微生物治疗方法。尽管单个病原体所传递的毒素各不相同，但组装和部署III型分泌系统的基本原理是很保守的，并且代表了潜在的治疗靶点。为了设计有效的策略来禁用这个毒力因子，更好地了解它的结构

DESCRIPTION (provided by applicant): Type III secretion is a method of delivering toxic proteins directly into host cells via a molecular syringe- like structure, which is employed by many Gram-negative pathogens. Type III secretion systems contribute to an incredibly diverse range of diseases in humans including bubonic plague, food-borne illnesses, and cystic fibrosis. The millions of cases each year of food-borne illnesses alone have a staggering impact on our health care system. Add to that the increasingly deadly potential from emerging antibiotic resistance, and the need for new antimicrobial treatments is urgent. Though the toxins that are delivered by individual pathogens vary, the fundamentals of assembling and deploying type III secretion systems are well conserved and represent a potential therapeutic target. To design effective strategies that disable this virulence factor, a better understanding of its architecture and regulation is necessary. Type III secretion systems are generally only made and activated during infection of a susceptible host. During infection, bacteria assemble the needle-like structure, but toxins are not secreted through it until contact with a host cell is made. How do bacteria activate the secretion system upon cell contact? Just as important, how do they shut if off? These questions are fundamental to our understanding of the system, and these questions are the focus of this proposal. We study the Yersinia type III secretion system because it is a well-established model system. Our novel tools and approaches have revealed new insight into the heart of type III secretion regulation. In particular, we have found that YopK, a regulator of type III secretion, has two distinct regulatory functions and works at both ends of the "needle". I appears that YopK is an integral component of an ON/OFF switch for secretion. From the bacterial side, YopK helps to monitor which proteins are secreted through the needle. YopK itself is secreted and once inside the host cell, YopK interacts with the end of the needle complex to shut down secretion of the toxins. In this proposal we will investigate how YopK performs these activities. Using a combination of genetics, cell biology, and biochemistry we will determine how YopK coordinates with other known secretion regulators on both sides of the needle. We will also identify and characterize the functional domains within YopK that mediate its activities. Collectively, our work will reveal fundamental mechanisms controlling this important virulence weapon. If we understand the nature of the ON/OFF switch, we can design a new type of broad-spectrum drug that targets a vital secretion system in a wide range of bacteria.