Identifying Regulation of Cell Cycle Gene Networks in E*

描述（由申请人提供）：本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资助R 01 AI 53724的扩展。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，寄生虫迅速增殖，然后可能导致结肠疾病。我们感兴趣的是确定调节这种原生寄生虫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。本研究的目的是通过基因芯片技术来鉴定新的阿米巴毒力因子。FIRCA补充是由Anuradha Lohia教授撰写的，他一直在研究溶组织内阿米巴细胞周期进程的调节。 阿努拉达在印度的实验室的结果表明，在E。溶组织菌基因组复制可能在没有细胞分裂的情况下发生，并且单个细胞核在增殖期可能含有几种基因组内容物（Gangopadhyay等人，1997 a; Das和Lohia，2002）。因此，与大多数真核生物不同，E.溶组织菌似乎不受已知的检查点监视机制控制。因此，在大肠杆菌中鉴定调控细胞周期进程的基因的全局谱是很重要的。溶组织剂在这个提议中，我们计划使用dsRNA干扰（Kaur和Lohia，2004）来下调细胞周期同源物- Eh Cdks和细胞周期蛋白，然后使用微阵列进行转录谱分析，以鉴定大肠杆菌中由CDK和细胞周期蛋白控制的基因组。溶组织剂这项工作对确定大肠杆菌细胞周期控制的新方面具有重要意义。溶组织剂此外，该项目是一个理想的合作环境，其主要目标是向印度的Lohia实验室转让技术（使用和分析微阵列数据）。阵列作为后基因组工具是强大的，但高度专业化，因此Lohia小组对该应用程序的“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.