Association of the histone-like protein HBsu with the nucleoid of Bacillus subtilis
Association of the histone-like protein HBsu with the nucleoid of Bacillus subtilis
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组蛋白样蛋白 HBsu 与枯草芽孢杆菌核的关联
DOI：
10.1128/jb.179.6.2060-2064.1997
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发表时间：
1997
影响因子：
3.2
通讯作者：
M. Marahiel
中科院分区：
文献类型：
--
作者：
Petra Köhler;M. Marahiel
为了研究枯草芽孢杆菌（Bacillussubtilis）的类组蛋白（HBsu）在类核结构中的生理作用，构建了与维多利亚水母（Aequoreavictoria）的绿色荧光蛋白（GFP）的融合体。这种纯化的融合蛋白，HBsuGFP，表现出三倍减少的亲和力，DNA相比，未经修饰的HBsu，然而，在凝胶迁移率变化实验HBsuGFP DNA结合大大增强，在存在低HBsu浓度。HBsu的额外产生对a B的延迟生长也有积极影响。枯草杆菌菌株PK 9 C8，其仅表达hbs-gfp（编码HBsuGFP）。HBsu似乎不仅影响生长，而且影响类核结构，通过DNA染色和荧光显微镜监测。在没有HBsu产生的情况下，菌株PK 9 C8显示出与HBsuGFP相关的松弛的类核结构。然而，当HBsu合成被诱导时，与融合蛋白的荧光一致的高度紧凑的类核结构被可视化。这为HBsu在DNA包装中的体内结合及其对细胞生长的影响提供了第一个证据。
To investigate the physiological role of the essential histone-like protein of Bacillus subtilis (HBsu) in the nucleoid structure, a fusion to the green fluorescent protein (GFP) of Aequorea victoria was constructed. This purified fusion protein, HBsuGFP, showed a threefold-reduced affinity to DNA compared to unmodified HBsu; however, in gel mobility shift experiments HBsuGFP DNA-binding was greatly enhanced in the presence of low HBsu concentrations. Additional production of HBsu also had a positive effect on the retarded growth of a B. subtilis strain, PK9C8, which expresses only hbs-gfp (encoding HBsuGFP). HBsu seemed to influence not only growth but also nucleoid structure, as monitored by DNA staining and fluorescence microscopy. Without HBsu production, strain PK9C8 showed a relaxed nucleoid structure associated with HBsuGFP. However, a highly compact nucleoid structure that coincides with the fluorescence of the fusion protein was visualized when HBsu synthesis was induced. This provides the first evidence for in vivo association of HBsu in DNA packaging and its consequence on cell growth.