Enhancing Undergraduate Research: Genome Data Mining and Laboratory Experimentation to Reveal the Fungal Agglutinin-Like Sequence Family

项目摘要 口咽念珠菌病(OPC)仍然是艾滋病毒感染者的常见病，尽管 抗逆转录病毒治疗的可用性。这项工作的长期目标是开发新的口腔治疗方法 HIV患者中的真菌疾病。治疗必须对白色念珠菌有效，最 OPC的常见病因，以及其他致病念珠菌物种。因为真菌细胞与 口腔粘膜是OPC所必需的，干扰真菌黏附将消除定植和 随后的疾病。白念珠菌凝集素样序列(Als)蛋白在 真菌细胞与口腔表面的粘连。AlS结构的最新结晶学解 粘附域揭示了负责粘附性相互作用的多肽结合空腔。 白色念珠菌和口腔上皮细胞之间的关系。因为ALS蛋白在其他致病菌中发现 引起OPC的假丝酵母菌，它们是抗粘连疗法开发的主要靶点。 然而，已发表的这些物种基因组中的ALS序列被错误地组装，因此 真菌ALS家族的组成和变异性尚不清楚。这种情况与此类似。 在白念珠菌基因组序列组装过程中出现的问题，可以补救 通过实验室实验确定每个ALS基因及其独特的物理位点。这个项目 将获得致病念珠菌中ALS基因的准确DNA序列，这些致病念珠菌会导致 OPC。基因表达分析将表明哪些编码蛋白可能具有主要的存在 在真菌细胞表面。候选蛋白质将被选择用于后续的结构分析和 设计对多种真菌有效的粘附性抑制剂。这个地区的建议书 将有来自伊利诺伊大学和密利金大学的本科生参加。学生们将 学习基于计算机的基因组数据分析，并获得有价值的分子生物学实验室技能。 该项目将改善两个机构的研究环境，并克服以下关键障碍 ALS类抗粘连疗法的研究进展。

Project Summary Oropharyngeal candidiasis (OPC) is still a common disease of HIV-infected people, despite the availability of antiretroviral therapy. The long-term goal of this work is to develop new treatments for oral fungal disease in HIV patients. Treatments must be effective against Candida albicans, the most common cause of OPC, and also other pathogenic Candida species. Because adhesion of fungal cells to the oral mucosa is required for OPC, interfering with fungal adhesion would eliminate colonization and subsequent disease. The agglutinin-like sequence (Als) proteins of C. albicans play a key role in adhesion of fungal cells to oral surfaces. Recent crystallographic solution of the structure of the Als adhesive domain revealed a peptide-binding cavity that is responsible for the adhesive interaction between C. albicans and oral epithelial cells. Because Als proteins are found in other pathogenic Candida species that cause OPC, they are prime targets for development of anti-adhesive therapies. However, ALS sequences in published genomes for these species are incorrectly assembled so the composition of and variability in the fungal ALS family remain unknown. This situation echoes similar problems that occurred during assembly of the C. albicans genome sequence and can be remedied through laboratory experimentation to define each ALS gene and its unique physical locus. This project will derive accurate DNA sequences for the ALS genes in the pathogenic Candida species that cause OPC. Gene expression analysis will indicate which encoded proteins are likely to have a major presence on the fungal cell surface. Candidate proteins will be selected for subsequent structural analyses and design of adhesive inhibitors that will be effective against multiple fungal species. This AREA proposal will involve undergraduate students from the University of Illinois and Millikin University. Students will learn computer-based analysis of genome data and gain valuable molecular biology laboratory skills. The project will enhance the research environment of both institutions and overcome a critical barrier to progress in the development of Als-based anti-adhesive therapies.