Identification and characterization of programmed cell death markers in bacterial models.
Identification and characterization of programmed cell death markers in bacterial models.
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DOI：
10.1007/978-1-62703-383-1_11
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发表时间：
2013-01-01
期刊：
影响因子：
--
通讯作者：
Winkler, Jonathan A
中科院分区：
文献类型：
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作者：
Dwyer, Daniel J;Winkler, Jonathan A
在面临末端应激的真核生物中，基因编码的细胞死亡途径的激活是核心细胞过程和关键生物分子功能修饰的根本变化的基础。这些生理变化在细胞程序性死亡过程中表现为表型特征，也是启动的特定死亡模式的标志。越来越多的工作表明，原核生物也能够表现出细胞程序性死亡的特征，尽管没有在高等生物体中控制这些事件的多个、紧密的调节层。本章描述了在真核模型中用于检测细胞程序性死亡特征的方法和材料是如何转移到原核模型的。特别是，我们描述了这些方法在研究抗生素后对细菌的影响方面的适用性，特别是由药物分子和靶点的相互作用引起的生化变化，包括氧化应激，伴随并确保细胞死亡。我们特别讨论了检测DNA断裂、染色体凝聚、磷脂酰丝氨酸暴露、膜去极化和caspase底物多肽结合的技术，从而为研究细菌中这些生理事件的进化提供了一个起点。
In eukaryotic organisms facing terminal stress, activation of genetically encoded cell death pathways underlies fundamental changes in core cellular processes and functional modification of critical biomolecules. These physiological alterations manifest themselves as phenotypic hallmarks during programmed cell death, and are markers of the particular mode of death initiated. A growing volume of work has illustrated that prokaryotes too are capable of exhibiting hallmarks of programmed cell death, albeit without the multiple, tight regulatory layers which control these events in higher order organisms.This chapter describes how methods and materials which have been used to assay for hallmarks of programmed cell death in eukaryotic models are transferrable to prokaryotic models. In particular, we describe the applicability of these methods to the study of post-antibiotic effects on bacteria, notably the biochemical changes induced by the interaction of drug molecules and targets, including oxidative stress, that accompany and ensure cell death. Specifically we discuss techniques for detecting DNA fragmentation, chromosomal condensation, phosphatidylserine exposure, membrane depolarization, and caspase substrate peptide binding, thereby providing a launchpoint for the study of the evolution of these physiological events in bacteria.