NOT1 in stress tolerance of Cryptococcus neoformans

NOT1在新型隐球菌应激耐受中的作用

• 批准号: 6915062
• 负责人: JOHN C PANEPINTO
• 金额: $ 4.99万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6915062
• 关键词: Cryptococcus neoformans; environmental adaptation; fungal genetics; gene environment interaction; gene expression; gene expression profiling; genetic regulation; genetic regulatory element; laboratory mouse; messenger RNA; miscellaneous oxidoreductase; mutant; northern blottings; nucleic acid metabolism; phenotype; postdoctoral investigator; stress; virulence

DESCRIPTION (provided by applicant): Cryptococcus neoformans is a major cause of morbidity and mortality in immunocompromised patients, especially in those afflicted with HIV/AIDS. The overall objective of this proposal is to test the hypothesis that NOT1 is a regulator of stress tolerance and virulence in C. neoformans. A homolog of S. cerevisiae NOT1, a global regulator of nutrient- and stress-sensitive transcription, was identified as an overexpressed transcript in the avirulent, stress-sensitive VAD1 mutant. Specific Aim 1 will define the role of NOT1 mRNA accumulation in stress tolerance and laccase expression in the delta vadl mutant and in wild type C. neoformans. This will be accomplished by profiling NOT1 expression in response to stress using northern blot, assessing the phenotypic consequences of NOT1 overexpression on the wild type, and assessing the effects of NOT1 mRNA suppression in the delta vadl mutant. Specific Aim 2 will determine the role of mRNh synthesis and degradation in the regulation of NOT1 mRNA abundance by separating the two processes experimentally, and define cis elements in the 5' and 3' flanking regions that influence its regulation. Exploring the NOT1 dependent stress response may reveal novel virulence factors in C. neoformans.

描述（由申请人提供）：隐孢子虫新生怪是免疫功能低下患者发病率和死亡率的主要原因，尤其是患有HIV/AIDS的患者。该提议的总体目的是检验假设，即Not1是新生甲虫中胁迫耐受性和毒力的调节剂。酿酒酵母Not1的同源物是养分和应激敏感性转录的全球调节剂，被确定为无抗应力的VAD1突变体中过表达的转录本。具体的目标1将定义NOT1 mRNA积累在胁迫耐受性和漆酶表达中的作用，并在野生型C. Neoformans中的作用。这将通过使用Northern blot响应压力来响应压力来实现这一点，从而评估NOT1过表达对野生型的表型后果，并评估Delta VADL突变体中Not1 mRNA抑制的影响。具体目标2将通过通过实验分开这两个过程，并在5'和3'侧翼区域中定义了影响其调节的5'和3'的CIS元素，从而确定MRNH合成和降解在调节Not1 mRNA丰度中的作用。探索NOT1依赖性应力反应可能揭示了Neoformans C. Neoformans中的新型毒力因子。