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MOLECULAR BIOLOGY OF CRYPTOCOCCUS NEOFORMANS

新型隐球菌的分子生物学

基本信息

批准号：
2003596
负责人：
JEFFREY C EDMAN
金额：
$ 12.38万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
1990
资助国家：
美国
起止时间：
1990-07-01 至 1997-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2003596
关键词：
Cryptococcus neoformans cryptococcosis fungal genetics genetic manipulation genetic regulatory element mass spectrometry molecular biology molecular cloning morphology nucleic acid sequence pheromone transcription factor virulence

项目摘要

DESCRIPTION: (Adapted from Applicant's Abstract). Cryptococcus neoformans remains a significant cause of morbidity and mortality in patients with AIDS and other immunosuppressive states. Despite advances in the treatment of cryptococcal meningitis, there is still no curative treatment in AIDS patients. In the past five years, there has been significant advances in the molecular biology of C. neoformans. Over a dozen genes have been cloned and characterized, transformation systems have been developed, and pioneering studies on virulence-related genes have been described. Overall, these studies have shown that not only is this organism an important field of study because of its relevance to human health, but it provides an ideal model system of fungal pathogenesis. The focus of the Edman laboratory, which has been an understanding of the mechanisms of dimorphism in C. neoformans, the characterization of the mating type loci, and the role of the MAT alpha locus in promoting virulence, will be continued through studies described in this application. Recent observations by Dr. Edman have demonstrated a true yeast to hyphae dimorphic transition in C. neoformans wild type, haploid, MAT alpha cells. The control of this transition will be characterized by the isolation of mutants in this pathway, cloning the genes defined by these mutants, and morphologic assessment of the hyphae produced in mutant and wild type strains. The mating type loci of C. neoformans will be characterized by transcript analysis, DNA sequence, and gene disruption. While there are over a dozen genes within these complex loci, two MATalpha genes that have already been isolated, the pheromone gene and a homologue of the S. cerevisiae PRT1 gene, will receive initial scrutiny. The mature pheromone will be characterized by purification from overexpressing cells and its structure determined by mass spectrometry. Cis- and trans-acting factors responsible for pheromone gene regulation will be identified, isolated and characterized. These studies will be complemented by the continuing isolation of C. neoformans mutants that are unable to form filaments after mating. MATalpha deletion and MATalpha::MATalpha replacement strains will be constructed to formally test the hypothesis that there are virulence determinants within the MATalpha locus. These strains should provide new insights into the mechanisms of pathogenesis of C. neoformans.

描述：(改编自申请人摘要)。隐球菌新生杆菌仍然是#年发病率和死亡率的重要原因。艾滋病和其他免疫抑制状态的患者。尽管取得了进步在治疗隐球菌性脑膜炎方面，目前还没有根治方法艾滋病患者的治疗。在过去的五年里，新生葡萄球菌分子生物学研究的重大进展。超过一年十几个基因已经被克隆和鉴定，转化系统已经开展了对毒力相关基因的开创性研究都被描述过。总体而言，这些研究表明，不仅是这种有机体是一个重要的研究领域，因为它与人类健康，但它提供了一个理想的真菌模型系统发病机制。埃德曼实验室的重点一直是对新生隐孢子虫的二型性机制的认识交配型基因座的特征和MATα基因的作用在提高毒力方面，将通过描述的研究继续进行在此应用程序中。埃德曼博士最近的观察表明新生隐孢子虫野生型中真正的酵母向菌丝二型转变，单倍体，垫阿尔法细胞。对这一过渡的控制将是以分离这一途径中的突变体为特征的，克隆这些突变体所定义的基因和菌丝的形态鉴定由突变株和野生型菌株生产。交配型基因座。新生杆菌将通过转录本分析，DNA序列，和基因破坏。虽然这些基因中有十几个复杂的基因座，已经分离出的两个MATAlpha基因，信息素基因和酿酒酵母PrT1基因的同源物，将接受初步审查。成熟的信息素的特征是高效表达细胞的纯化及其结构的确定质谱学。顺式和反式作用因素导致信息素基因的调控将被识别、分离和表征。这些研究的补充将是对C. 在交配后不能形成细丝的新生物突变体。 MATAlpha缺失和MATAlpha：：MATAlpha替换菌株将是被构造为正式测试存在毒力的假设 MATAlpha基因座内的决定因素。这些菌株应该会提供新的对新生葡萄球菌致病机制的洞察。