Identifying Regulation of Cell Cycle Gene Networks in E*

描述(由申请人提供)：这项研究将主要在印度加尔各答的Bose研究所与Anuradha Lohia合作完成，作为NIH赠款R01AI 53724的延伸。人类病原体溶组织内阿米巴是引起阿米巴痢疾和肝脓肿的原因。人体肠道感染后，寄生虫很快就会繁殖繁殖，这可能会导致结肠疾病。我们感兴趣的是在这种原生寄生虫中识别调节细胞增殖的分子因子，以努力设计新的策略来预防阿米巴病。父母建议的目标是通过基于微阵列的转录图谱来识别新的阿米巴毒力因子。FIRCA附录是由Anuradha Lohia教授撰写的，他一直在研究溶组织内阿米巴的细胞周期进程调节。 Anuradha在印度的实验室的结果表明，在溶组织埃希氏菌中，在增殖期可能发生基因组复制而不进行细胞分裂，并且单个核可能包含几个基因组内容(Gangopadhyay等人，1997a；Das和Lohia，2002年)。因此，与大多数真核生物不同，溶组织棘球绦虫的细胞周期似乎不受已知的检查点监视机制的控制。因此，重要的是要确定全球范围内调控细胞周期进程的基因在组织裂解杆菌中的分布。在这项建议中，我们计划使用dsRNA干扰(Kaur和Lohia，2004)下调细胞周期同源物-Eh CDKs和Cyclins，然后使用微阵列进行转录谱分析，以确定由CDKs和Cyclins控制的基因组。这项工作对鉴定溶组织埃希氏菌细胞周期调控的新方面具有重要意义。此外，该项目是一个理想的合作环境，主要目标是向印度的洛伊亚实验室转让技术(使用和分析微阵列数据)。阵列作为一种后基因组工具是强大而高度专业化的，因此Lohia的团队对这一应用程序的“动手”使用是这项工作的重要组成部分。

DESCRIPTION (provided by applicant): This research will be done primarily at the Bose Institute in Calcutta, India in collaboration with Anuradha Lohia as an extension of NIH grant R01AI 53724. The human pathogen Entamoeba histolytica is responsible for causing amoebic dysentery and liver abscesses. Infection of the human gut is quickly followed by proliferation and multiplication of the parasite, which may then lead to colonic disease. We are interested in identifying molecular factors regulating cell proliferation in this protist parasite, in an effort to devise new strategies to prevent amoebiasis. The goal of the parent proposal is to identify novel amoebic virulence factors by microarray based transcriptional profiling. The FIRCA supplement is written with Prof. Anuradha Lohia who has been studying the regulation of cell cycle progression Entamoeba histolytica. Results from Anuradha's laboratory in India have shown that in E. histolytica genome reduplication may occur without cell division and single nuclei may contain several genome contents during the proliferative phase (Gangopadhyay et al, 1997a; Das and Lohia, 2002). Therefore, unlike most eukaryotes the cell cycle of E. histolytica does not appear to be controlled by known checkpoint surveillance mechanisms. It would be important therefore to identify a global profile of genes that regulate cell cycle progression in E. histolytica. In this proposal we plan to use dsRNA interference (Kaur and Lohia, 2004) to down-regulate cell cycle homologues- Eh Cdks and cyclins, followed by transcriptional profiling using microarrays in order to identify groups of genes controlled by CDKs and cyclins in E. histolytica. This work has important implications for identifying novel aspects of cell cycle control in E. histolytica. Additionally, this project is an ideal collaborative environment in which a main goal is technology transfer (use and analysis of microarray data) to the Lohia lab in India. Arrays as a post-genomic tools are powerful yet highly specialized, thus the "hands-on" use of this application by Lohia's group is an important component of this work.