STRUCTURAL BIOLOGY OF BACTERIAL PATHOGENS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 这项研究的目的是研究1)一组新的DNA结合蛋白，称为α/β型小分子酸溶蛋白(SASP)如何保护休眠芽胞中的DNA免受各种损害剂的伤害，以及2)这些蛋白如何在孢子生长过程中随后被移除，最终导致3)染色体复制。这些研究将包括对α/β型SASP及其与DNA的络合物的生化和生物物理性质的阐明，对在孢子生长过程中启动SASP降解的孢子蛋白酶GPR的表征，以及对负责染色体DNA复制的新型细菌聚合酶POLE和PolC(apo和与DNA模板：引物的复合体)的表征。拟议研究的一个主要目标是所有这些成分的表征(机理和功能)，包括用X射线结晶学阐明它们的三维结构。成功地实现这一目标将在分子水平上理解休眠孢子中DNA保护的机制，导致孢子萌发和生长过程中DNA保护丧失的机制，以及后来在营养细胞中复制DNA的机制。

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. The objectives of the research are to investigate 1) the mechanisms whereby a group of novel DNA-binding proteins, termed alpha/beta-type small, acid-soluble proteins (SASP) protect DNA in dormant spores of Bacillus species against a variety of damaging agents, and 2) how these proteins are subsequently removed during spore outgrowth, ultimately leading to 3) chromosomal replication. The studies will involve biochemical and biophysical elucidation of the properties of the alpha/beta-type SASP and the complexes of these proteins with DNA, the characterization of the spore protease, termed Gpr, that initiates SASP degradation during spore outgrowth, as well as characterization of PolE and PolC (apo and complexes with DNA template:primers) novel bacterial polymerases responsible for chromosomal DNA replication. A major goal of the proposed research is the characterization (mechanistic and functional) of all of these components including elucidation of their three-dimensional structures by x-ray crystallography. Success in achieving this goal will lead to an understanding at the molecular level of the mechanisms of DNA protection in dormant spores, the mechanisms responsible for the loss of this DNA protection during spore germination and outgrowth, and later mechanism of its replication in vegetative cells.