The Role of Unc119 in T Cell Antigen Receptor Signaling1

描述（改编自摘要）：新型隐球菌是免疫功能低下患者的主要病原体，在大约6%的HIV阳性个体中引起危及生命的脑膜脑炎。体外黑色素的产生与C.新人类在上一个资助期内，C.新形式疟原虫的研究确立了CNLAC 1基因在毒力中的重要性，并且在感染小鼠的脑中鉴定了漆酶依赖性儿茶酚胺氧化产物。此外，一个复杂的模式的调控DNA结合位点的CNLAC 1基因上游已被确定超过被确定与一个不寻常的Sp1增强子位点。在其他毒力基因如Cap 64和Cap 59中发现了Sp1共有序列，表明毒力的共同调节。待检验的假设是CNLAC 1的分子调节剂控制新生隐球菌的毒力。拟开展的研究的目的是鉴定和表征参与漆酶表达的基因，以确定它们在毒力中的作用。目的1提出了确定和表征重要的调控DNA结合位点的上游区域的CNLAC 1。该计划是使用pVEW启动子质粒和电迁移率变动分析来确定在葡萄糖阻遏和去阻遏以及37 ℃的宿主温度条件下的显著增强子和阻遏子区域。目的2提出鉴定和表征参与CNLAC 1抑制的转录增强的基因，并确定它们在毒力中的作用。基因将通过与CNLAC 1 DNA结合位点预测的基因和来自漆酶缺陷插入突变体的基因的同源性来克隆。对于每个基因，计划是产生敲除和野生型互补的C.并测试该基因对漆酶产生、荚膜形成、分泌的甘露糖蛋白、脲酶活性和毒力的影响。目的3从DL-1突变体中鉴定和鉴定参与漆酶转录后修饰的基因。最初，计划是补充漆酶分泌的液泡H+-ATP酶，并评估CNLAC 1和CNVPH 1可能的共同调节。预计这些研究将为深入了解C.新型隐球菌，其可用于提供治疗和控制隐球菌病的新方法。

Description (Adapted from Abstract): Cryptococcus neoformans is a major pathogen in immunocompromised patients, causing life-threatening meningoencephalitis in approximately 6% of HIV positive individuals. In vitro melanin production has classically been associated with virulence in C. neoformans. During the previous funding period congenic knockout strains of C. neoformans establish the importance of the CNLAC1 gene in virulence and laccase-dependent catecholamine oxidation products have been identified in the brain of infected mice. Furthermore, a complex pattern of regulatory DNA-binding sites upstream of the CNLAC1 gene have been identified over been identified together with an unusual Sp1 enhancer site. Sp1 consensus sequences have been found in the other virulence genes such as Cap64 and Cap59 suggesting co-regulation of virulence. The hypothesis to be examined is that molecular regulators of CNLAC1 control virulence of Cryptococcus neoformans. The objectives of the proposed research are to identify and characterize genes involved in laccase expression to determine their role in virulence. Aim 1 proposes to identify and characterize important regulatory DNA-binding sites in the upstream region of CNLAC1. The plan is to use pVEW promoter plasmid and electromobility shift assays to determine significant enhancer and repressor regions under conditions of glucose repression and derepression as well as the host temperature of 37 C. Aim 2 proposes to identify and characterize genes involved in transcriptional enhancement of repression of CNLAC1 and determine their role in virulence. Genes will be cloned by homology to genes predicted by CNLAC1 DNA-binding sites and from laccase-deficient insertional mutants. For each gene the plan is to produce knockout and wild-type complemented strains of C. neoformans and test the effects of the gene on laccase production, capsule formation, secreted manno-proteins, urease activity and virulence. Aim 3 will identify and characterize genes involved in post-transcriptional modification of laccase from DL-1 mutants. Initially, the plans are to complement a vacuolar H+-ATPase on laccase secretion and assess for possible co-regulation of CNLAC1 and CNVPH1. It is anticipated that these studies will provide insights into regulation and virulence factors in C. neoformans that may be used to provide novel approaches to the treatment and control of cryptococcosis.