RESPONSE REGULATOR GENE (CASSK1) OF CANDIDA ALBICANS

描述（改编自申请人摘要）：白色念珠菌是最常见的 艾滋病患者常见的真菌病原体。感染最常涉及 口腔、食道或阴道的粘膜表面。尽管有 三唑类药物作为抗真菌药物的出现， 念珠菌病是复杂的耐药C。白色念珠菌和 非白色念珠菌种属。合适的替代品范围有限 这就需要确定新的目标， 开发抗真菌药物。我们的方法是针对非哺乳动物 磷蛋白是重要的信号转导途径的组成部分 白色念珠菌为此，我们已经确定了一个2组分，组氨酸激酶 （CaHKI）和应答调节基因（CaSSKl），这两者对于调节细胞增殖和分化是必需的。 生物体的毒力，因为每个基因中的敲除菌株是无毒的 与亲本、杂合子和基因重建菌株相比。的 这项提案的重点将放在CaSSKl上，这是第一个分离的响应调节器 梭白色念珠菌我们强调这个基因的理由是双重的。一是其 缺失导致无毒。第二，该基因的一个同源物， 粟酒裂殖酵母为许多 应激条件，包括氧化，高温和蛋白质 抑制剂应激这项建议有三个具体目标。在特定 目标1：体外研究将解决氧化应激问题。表达将 通过北方分析和竞争性PCR平行测量， 翻译后研究（磷酸化）。在具体目标2中， CaSSK1缺失菌株（及其单拷贝配偶）的基因组将在 念珠菌病的阴道和口腔模型。具体目标3将侧重于 用减法识别Casskip下游效应器 杂交和酵母双杂交系统。这一长期目标 研究是为了扩大抗C的数量。白色念珠菌目标和信息 这将导致开发高通量检测方法， 潜在的抑制剂

DESCRIPTION (Adapted from applicant's abstract): Candida albicans is the most common fungal pathogen of AIDS patients. The infection most often involves the mucosal surfaces of the oral cavity, esophagus or vagina. In spite of the advent of triazoles as anti-fungals, management of AIDS patients with candidiasis is complicated by the development of drug-resistant C. albicans and non-albicans species. The spectrum of suitable alternatives is limited underlying the need to identify new targets, which can be exploited in the development of anti-fungals. Our approach has been to target non-mammalian phosphoproteins that are components of important signal transduction pathways of C albicans. To this end, we have identified a 2-component, histidine kinase (CaHKI) and response regulator genes (CaSSKl), both of which are essential for virulence of the organism, since knock-out strains in each gene were avirulent in comparison to parental, heterozygote and gene-reconstituted strains. The focus of this proposal will be on CaSSKl, the first response regulator isolated from C. albicans. Our reasons for emphasizing this gene are 2-fold. First, its absence results in avirulence. Second, a homologue of this gene in Schizosaccharomyces pombe provides essential adaptation functions for a number of stress conditions, including oxidative, high temperature and protein inhibitor stress. There are three specific aims in this proposal. In specific aim 1, in vitro studies will address oxidative stress. Expression will be measured by Northern analysis and competitive PCR in parallel with post-translational studies (phosphorylation). In specific aim 2, the virulence of CaSSKl-deleted strains (and their single-copy mates) will be evaluated in vaginal and oral models of candidiasis. Specific aim 3 will focus upon the identification of down-stream effectors of Casskip using subtractive hybridization and the yeast 2-hybrid system. The long-term objectives of this research are to expand the number of anti-C. albicans targets and information which will lead to the development of high-throughout assays for evaluating potential inhibitors.