SCHISTOSOME NUCLEAR PROTEIN FUNCTION AND REGULATION

血吸虫核蛋白的功能和调节

• 批准号: 2005822
• 负责人: James W Tracy
• 金额: $ 19.22万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2005822
• 关键词: Escherichia coli; Saccharomyces cerevisiae; Schistosoma mansoni; cell cycle; developmental genetics; fluorescence microscopy; genetic library; immunocytochemistry; laboratory mouse; life cycle; messenger RNA; northern blottings; nucleoproteins; protein sequence; protein structure function; tissue /cell culture; yeast two hybrid system

Schistosomiasis remains the most prevalent trematode infection of humans afflicting over 200 million people. There is no vaccine and praziquantel is the only broad-spectrum antischistosomal drug available. The best long- term defense against the appearance and propagation of drug-resistant parasites is continued basic research directed toward understanding the biology of schistosomes as it relates to the identification of novel biochemical and cellular targets for rational drug design. Most cells of adult Schistosoma mansoni are quiescent. The principal exceptions are the reproductive organs, especially the female vitelline glands. Despite of the importance of egg production in pathogenesis and transmission of schistosomiasis, little is known about schistosome cell proliferation. Understanding the role of nuclear proteins in this process could point to alternative approaches for controlling the infection by interfering with parasite development and reproduction. This proposal focuses on one such protein, SMAK (schistosome homologue of the yeast nuclear protein MAK16), as a prototype. Not only do SMAK and MAK16 share significant amino acid sequence similarity, but SMAK can also complement the makl6-1 mutation in yeast. This proposal is designed to investigate the biochemical function of SMAK and to begin to explore its regulation. There are four specific aims: l) To delineate the tissue, cell and subcellular distribution of SMAK in adult S.mansoni using immunocytochemical techniques and specific anti-SMAK antibodies; 2) To determine whether SMAK expression is develop mentally regulated by measuring the level of SMAK messenger RNA in various schistosome stages; 3) To identify proteins that interact with SMAK using a yeast two-hybrid screen to distinguish between a role in cell cycle progression and 60S ribosome subunit biogenesis: 4) To characterize protein sequence motifs that are hypothesized to be involved in the biochemical function and/or regulation of SMAK. These include: a) a novel zinc binding domain: b) two putative nuclear localization sequences: and c) consensus sequences for phosphorylation of SMAK by protein kinase CK2.

血吸虫病仍然是人类最常见的吸虫感染 影响超过2亿人。没有疫苗和吡喹酮 是唯一可用的广谱抗血吸虫药物。最好的长—— 针对耐药性出现和传播的术语防御 寄生虫是持续的基础研究，旨在了解 血吸虫生物学，因为它与新型血吸虫的鉴定有关 合理药物设计的生化和细胞靶点。 成年曼氏血吸虫的大多数细胞是静止的。校长 生殖器官除外，尤其是雌性卵黄 腺体。尽管产蛋在发病机制中的重要性和 血吸虫病的传播途径，对血吸虫细胞知之甚少 增殖。了解核蛋白在此过程中的作用 可以指出控制感染的替代方法 干扰寄生虫的发育和繁殖。这个提议 重点关注这样一种蛋白质：SMAK（酵母的血吸虫同源物） 核蛋白 MAK16)，作为原型。 SMAK 和 MAK16 不仅共享 显着的氨基酸序列相似性，但 SMAK 也可以互补 酵母中的makl6-1突变。本提案旨在调查 SMAK的生化功能并开始探讨其调控。那里 有四个具体目标： l) 描绘组织、细胞和亚细胞 使用免疫细胞化学技术检测 SMAK 在曼氏血吸虫成虫中的分布 和特异性抗SMAK抗体； 2) 判断SMAK是否表达 通过测量 SMAK 信使 RNA 的水平来发展精神调节 处于不同的血吸虫阶段； 3) 鉴定与相互作用的蛋白质 SMAK 使用酵母双杂交筛选来区分细胞中的作用 循环进程和 60S 核糖体亚基生物发生：4) 表征 假设参与的蛋白质序列基序 SMAK 的生化功能和/或调节。其中包括：a) 小说 锌结合结构域：b) 两个推定的核定位序列：和 c) 蛋白激酶 CK2 磷酸化 SMAK 的共有序列。