SCHISTOSOME NUCLEAR PROTEIN FUNCTION AND REGULATION

血吸虫病仍然是人类最普遍的吸虫感染 超过2亿人受到影响没有疫苗和吡喹酮 是唯一可用的广谱抗组胺药。最好的长- 对耐药菌的出现和繁殖的长期防御 寄生虫是继续基础研究，旨在了解 与鉴定新的 合理药物设计的生化和细胞靶点。 曼氏血吸虫成虫的细胞大多数是静止的。校长 例外的是生殖器官，特别是雌性卵黄 腺体尽管产蛋在发病机制中的重要性， 血吸虫病的传播，对血吸虫病细胞的了解很少 增殖了解核蛋白在这一过程中的作用 可以指出控制感染的替代方法， 干扰寄生虫的发育和繁殖。这项建议 重点介绍了一种这样的蛋白质，SMAK（酵母的同源体 核蛋白MAK 16）作为原型。SMAK和MAK 16不仅共享 显著的氨基酸序列相似性，但SMAK也可以互补 酵母中的mak 16 -1突变。本提案旨在调查 SMAK的生化功能，并开始探索其调控。那里 有四个具体的目标：1）描绘组织，细胞和亚细胞 应用免疫细胞化学技术研究SMAK在曼氏血吸虫成虫中的分布 和特异性抗SMAK抗体; 2）为了确定SMAK表达是否 是通过测量SMAK信使RNA的水平来调节的 在不同的发育阶段; 3）为了鉴定与 SMAK使用酵母双杂交筛选来区分细胞中的作用 细胞周期进程和60 S核糖体亚基生物合成：4）表征 蛋白质序列基序，假设参与了 SMAK的生物化学功能和/或调节。其中包括：（1）小说 锌结合结构域：B）两个推定的核定位序列;和 c）通过蛋白激酶CK 2磷酸化SMAK的共有序列。

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.