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HIV感染者的常见疾病，尽管 提供抗逆转录病毒疗法。这项工作的长期目标是开发新的治疗口腔 艾滋病病毒感染者的真菌病。治疗必须对白色念珠菌有效， OPC的常见原因，以及其他致病性念珠菌属物种。因为真菌细胞粘附在 口腔粘膜是OPC所需的，干扰真菌粘附将消除定植， 后续疾病。C.白色念珠菌在 真菌细胞粘附于口腔表面。Als结构的最新晶体学解 粘附结构域揭示了负责粘附相互作用的肽结合腔 C.白色念珠菌和口腔上皮细胞。因为Als蛋白在其他致病性 念珠菌属引起OPC，它们是开发抗粘附疗法的主要目标。 然而，这些物种的已发表基因组中的ALS序列是错误组装的，因此， 真菌ALS家族的组成和变异性仍然未知。这种情况与类似的 在C.白念珠菌基因组序列，并可以补救 通过实验室实验来确定每个ALS基因及其独特的物理位点。这个项目 将推导出致病念珠菌中ALS基因的准确DNA序列， OPC。基因表达分析将表明哪些编码蛋白可能主要存在于 在真菌细胞表面。将选择候选蛋白用于随后的结构分析， 设计对多种真菌有效的粘附抑制剂。本区域提案 将涉及来自伊利诺伊大学和密立金大学的本科生。学生将 学习基于计算机的基因组数据分析，并获得宝贵的分子生物学实验室技能。 该项目将改善这两个机构的研究环境，并克服一个关键障碍， 在开发基于Als的抗粘连疗法方面取得进展。

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.