Glycosylation & Biogenesis of Streptococcal Fimbriae

描述（由申请人提供）：吴博士分离并鉴定了副猪链球菌的糖基化菌毛结构亚基Fap1分子，这是了解革兰氏阳性菌菌毛生物合成和粘附基础的重大突破。参与fap1糖基化的fap1样分子和基因不仅在口腔链球菌中高度保守，而且在肺炎链球菌和葡萄球菌中也存在。吴博士目前对Fap1糖基化的研究非常令人兴奋。很可能革兰氏阳性细菌中存在一种普遍的糖基化机制，这些研究的结果将适用于其他生物体。最近，他发现了Fap1糖基化所需的一个基因簇。他目前正在利用分子遗传学、结构生物学和生化方法确定Fap1糖基化的生物合成途径。在K22奖的支持下，吴博士将参加质谱和生物信息学的正式课程，并接受碳水化合物结构的质谱分析和蛋白质糖基化关键酶的生化分析的实践培训。K22的支持将为吴博士提供碳水化合物生物化学方面的新技能和知识，这将使他在链球菌黏附的糖基化研究中寻求新的途径。吴博士在确定原核糖基化和革兰氏阳性菌毛生物发生机制方面具有独特的地位，因为在过去的20年里，法夫斯-泰勒博士的实验室已经产生了明确的遗传工具。吴博士在分子遗传学方面的优势，加上碳水化合物生物化学特别是质谱分析和酶学方面的培训，将促进他在这个令人兴奋的研究领域的职业发展。这一建议的结果将有助于确定Fap1糖基化的生物合成途径。该途径中的新基因可能赋予微生物新的效应功能或帮助逃避宿主防御，这些策略是许多细菌病原体致病能力的重要组成部分。因此，阐明Fap1糖基化可能为理解糖基化在细菌发病机制中的作用提供一个框架。

DESCRIPTION (provided by applicant): Dr. Wu's isolation and characterization of the Fap1 molecule, a glycosylated fimbrial structural subunit of Streptococcus parasanguis, was a significant breakthrough in understanding the basic for fimbrial biosynthesis and adhesion in Gram-positive bacteria. Fap1-like molecules and genes involved in Fapl glycosylation are not only highly conserved across oral streptococci but are also present in Streptococcus pneumoniae and in the staophlococci. Dr. Wu's current research on the glycoslylation of Fap1 is extremely exciting. It is likely that a universal glycosylation machinery is present in Gram-positive bacteria and the findings from these studies would be applicable to other organisms. Recently, he identified a gene cluster that is required for the glycosylation of Fap1. He is currently in the process of determining the biosynthetic pathway by which Fap1 is glycosylated using molecular genetic, structural biology, and biochemical approaches. With the support of a K22 award, Dr. Wu will take formal courses in mass spectrometry and bioinformatics and receive hands-on training in Mass spectrometry analysis of the carbohydrate structure and biochemical analysis of key enzymes involved in protein glycosylation. The support of a K22 will provide Dr. Wu with new skills and knowledge in carbohydrate biochemistry that will position him to pursue new avenues in the study of glycosylation of streptococcql adhesions. Dr. Wu is in a unique position to determine the mechanisms of prokaryotic glycosylation and Gram-positive fimbrial biogenesis as the explict genetic tools have been generated in Dr. Fives-Taylor's laboratory over the last 20 years. Dr. Wu's strength in molecular genetics coupled with the training in carbohydrate biochemistry specifically in mass spectrometry analysis and enzymology will enhance his career development in this exciting research area. The results of this proposal will help define the biosynthetic pathway for Fap1 glycosylation. New genes in the pathway may endow microbes with new effector functions or aid in the evasion of host defense, strategies that are important components of disease-causing capabilities of a number of bacterial pathogens. Therefore, elucidation of Fap1 glycosylation may provide a framework for understanding the role of glycoslyation in bacterial pathogenesis.