CHARACTERIZATION OF PUTATIVE MAGNESIUM TRANSPORTING P-TYPE ATPASES

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 简介：P型ATP酶包括蛋白质的大超家族，存在于原核生物和真核生物中，其跨细胞膜运输无机阳离子和可能的其它底物。基于系统发育研究，它们被分为5个亚家族，称为P1-P5，根据序列相似性，跨膜组织和底物特异性进行分组。了解最少的P型ATP酶是P5亚家族的那些，其仅在真核生物中表达。目前建议的总体目标是获得有关小鼠中P5-ATP酶的基本信息，这对于最终了解其细胞生物学和生理功能至关重要。本研究的目的是：1）分离每个家族成员的全长cDNA克隆，并确定其组织分布和膜定位; 2）在适当的细胞系中过表达新的P型ATP酶，以评估其在细胞中的表达。 3）利用RNA干扰（RNAi）和基因靶向技术确定它们在体内的细胞和生理作用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Introduction: P-type ATPases comprise a large superfamily of proteins, present in both prokaryotes and eukaryotes, that transport inorganic cations and possibly other substrates across cell membranes. Based on phylogenetic studies they have been classified into 5 subfamilies, termed P1-P5, which group according to sequence similarities, transmembrane organization, and substrate specificity. The most poorly understood P-type ATPases are those of the P5 subfamily, which are expressed only in eukaryotes. The overall goal of the current proposal is to obtain basic information about the P5-ATPases in mice that will be essential for an eventual understanding of their cell biological and physiological functions. The aims of the current proposal are to 1) isolate full-length cDNA clones for each of the family members and determine their tissue distribution and membrane location, 2) overexpress the novel P-type ATPases in the appropriate cell lines in order to assess their i on specificity, and 3) determine their cellular and physiological roles in vivo using RNA interference (RNAi) and gene targeting technologies.