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Structural and Functional Diversity of alpha-Proteobacterial Acidocalcisomes/polyphosphate

α-变形菌酸钙体/多磷酸盐的结构和功能多样性

基本信息

批准号：
327837089
负责人：
Professor Dr. Dieter Jendrossek
金额：
--
依托单位：
Institut für Mikrobiologie (IMB)
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2016
资助国家：
德国
起止时间：
2015-12-31 至 2020-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/327837089?language=en
关键词：
Structural Functional Diversity alpha Proteobacterial

项目摘要

In the proposed project we will determine the structural and functional differences between bacterial acidocalcisomes (polyphosphate inclusions in a vacuolar proton pyro-phosphatase [V-H+-PPase, HppA]-driven acidic membrane-enclosed compartment) and non-membrane-enclosed polyphosphate granules of non-acidocalcisome-forming bacteria. We postulate that the presence of an hppA gene is indicative for the ability to form acidocalcisomes (all genome-sequenced alpha-proteobacteria have a hppA gene and the formation of acidocalcisomes in Agrobacterium tumefaciens and Rhodospirillum rubrum has been shown). We will isolate acidocalcisomes from A. tumefaciens (with membranes) and from Caulobacter crescentus (postulated to have membrane-enclosed acidocalcisomes). We will determine the elemental composition of acidocalcisomes, identify the acidocalcisome-associated proteins and determine the lipid-composition of the acidocalcisome membrane by in vitro and in vivo methods. This knowledge will enable us to compare bacterial acidocalcisomes with acidocalcisomes from eukaryotic unicellular counterparts (e. g. Trypanosomas cruzi) and with polyP granules from bacteria lacking a lipid membrane around polyphosphate granules. We will investigate the function of bacterial acidocalcisomes by a combination of growth and life imaging experiments and phenotype analysis of the wild type and generated mutant strains in which the key gene necessary for the formation of the acidic acidocalcisome compartment (V-H+-PPase, hppA) and/or the key gene of polyP synthesis (polyP kinase gene, ppk) are deleted in A. tumefaciens or C. crescentus. This project will considerably increase our knowledge on bacterial organelles.

在拟议的项目中，我们将确定细菌酸性钙体(液泡质子焦磷酸酶[V-H+-PPase，HppA]驱动的酸性膜封闭隔室中的聚磷酸盐包裹体)与非酸性钙体形成细菌的非膜包裹聚磷酸盐颗粒之间的结构和功能差异。我们推测，hppA基因的存在表明了形成酸性钙体的能力(所有基因组测序的α-蛋白细菌都有hppA基因，并且已经显示在根癌农杆菌和红色红螺菌中形成了酸性钙体)。我们将从根癌农杆菌(有膜)和新月杆菌(推测有膜包裹的酸性钙体)中分离出酸性钙体。我们将通过体外和体内方法测定酸性钙体的元素组成，鉴定酸性钙体相关蛋白，并测定酸性钙体膜的脂组成。这一知识将使我们能够比较细菌的酸性钙体与真核单细胞对应的酸性钙体(例如克氏锥虫)，以及与缺乏聚磷酸盐颗粒周围脂膜的细菌的息肉颗粒。我们将通过生长和生命成像实验以及野生型和突变株的表型分析相结合的方法来研究细菌酸性钙体的功能。在这些突变株中，酸性酸性钙体形成所必需的关键基因(V-H+-PPase，hppA)和/或息肉合成的关键基因(Polyp Kinase，PPK)在根癌农杆菌或新月弯曲霉中缺失。这个项目将大大增加我们对细菌细胞器的了解。