Stress-Responsive RNA Regulons in Cryptococcus neoformans

描述(申请人提供)：在世界上受艾滋病毒感染困扰的地区，致病真菌新生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胁迫耐受中的作用。我们的长期目标是系统地定位新生隐孢子菌维持胁迫耐受性和致病性所需的蛋白质-RNA相互作用。我们假设，这些RNA-蛋白质相互作用有助于新生葡萄球菌的适应性，使其对独立于核的基因表达具有敏捷性。在这个建议中，我们的重点是双管齐下的：首先，我们将鉴定与核糖体蛋白转录本中GGAUG顺式元件相互作用的RNA结合蛋白，该转录本在CCR4？突变体，并确定其在胁迫耐受性和致病性中的作用。其次，我们将确定新生葡萄球菌PUF家族的RNA结合蛋白在胁迫耐受性和致病性中的作用。对于GGAUG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互补，使我们能够组装它们各自的RNA调节子。未来的研究将建立在这项工作的基础上，并扩展到其他类别或RNA结合蛋白。最终，RNA-蛋白质相互作用的鉴定对新生芽孢杆菌的应激耐受性和致病性非常重要，这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.