Stress-Responsive RNA Regulons in Cryptococcus neoformans

描述（由申请人提供）：在世界上受HIV感染困扰的地区，致病性真菌新隐球菌已成为社区获得性脑膜炎的主要原因。我们之前的研究已经证明了转录后基因调控在应激耐受性中的作用。我们的长期目标是系统地绘制蛋白质- rna相互作用的图谱，这些相互作用是维持新生芽孢杆菌的耐受性和致病性所必需的。我们假设这些rna -蛋白质的相互作用有助于C. neoformans的适应性，赋予基因表达的灵活性独立于细胞核。在这个提案中，我们的重点是双管齐下：首先，我们将确定与在ccr4？突变，并确定其在胁迫耐受性和致病性中的作用。其次，我们将确定新生芽孢杆菌PUF家族RNA结合蛋白在胁迫耐受性和致病性中的作用。对于ggag结合蛋白和PUF蛋白，我们将确定与它们相互作用的转录本的补体，使我们能够组装它们各自的RNA调控。未来的研究将建立在这项工作的基础上，并扩展到其他种类的RNA结合蛋白。最终，鉴定rna -蛋白相互作用对新生芽孢杆菌的耐受性和致病性至关重要，将为我们提供新的基于小分子的治疗方法的特定靶点。

DESCRIPTION (provided by applicant): The pathogenic fungus Cryptococcus neoformans has emerged as the primary cause of community acquired meningitis in regions of the world plagued by HIV infection. Our previous studies have demonstrated a role for post-transcriptional gene regulation in stress tolerance. Our long term goal is to systematically map the protein-RNA interactions that C. neoformans requires for the maintenance of stress tolerance and pathogenicity. We hypothesize that these RNA-protein interactions contribute to the adaptability of C. neoformans, conferring agility to gene expression independent of the nucleus. In this proposal, our focus is two-pronged: First we will identify the RNA binding protein that is interacting with a GGAUG cis element in ribosomal protein transcripts found up-regulated in the ccr4? mutant, and determine its role in stress tolerance and pathogenicity. Second, we will determine the role of the C. neoformans PUF family of RNA binding proteins in stress tolerance and pathogenicity. For both the GGAUG-binding protein and the PUF proteins, we will determine the complement of transcripts that interact with each, allowing us to assemble their respective RNA regulons. Future studies will then build on this work and expand to additional classes or RNA binding proteins. Ultimately, the identification of RNA-protein interactions important for C. neoformans stress tolerance and pathogenicity will provide us with specific targets for novel small-molecule based therapeutics. PUBLIC HEALTH RELEVANCE: The long term goal of this project is to determine the RNA-protein interactions which C. neoformans requires for stress adaptation and pathogenesis. These studies will enhance our understanding of the molecular reprogramming that accompanies stress adaptation important pathogen. In defining these important RNA- protein interactions, we will determine those that will be viable targets for small-molecule therapeutics.