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核仁相关 Rok 蛋白差异影响枯草芽孢杆菌重组缺陷细胞的染色体转化。

Nucleoid-associated Rok differentially affects chromosomal transformation on Bacillus subtilis recombination-deficient cells.

机构信息

Department of Microbial Biotechnology, Centro Nacional de Biotecnología, CNB-CSIC, Madrid, 28049, Spain.

出版信息

Environ Microbiol. 2021 Jun;23(6):3318-3331. doi: 10.1111/1462-2920.15562.

Abstract

Rok, a Bacillus subtilis nucleoid-associated protein (NAP), negatively regulates competence development and silences xenogeneic genes. We show that rok inactivation increases rpoB482 natural intraspecies chromosomal transformation (CT) and plasmid transformation to a different extent. In ΔaddAB, ΔrecO, recF15, ΔrecU, ΔruvAB or rec cells intraspecies CT significantly increases, but the ΔrecD2 mutation reduces, and the ΔrecX, ΔradA or ΔdprA mutation further decreases CT in the Δrok context when compared to rok cells. These observations support the idea that rok inactivation, by altering the topology of the recipient DNA, differentially affects the integration of homologous DNA in rec-deficient strains, and in minor extent the competent subpopulation size. The impairment of other NAP (Hbsu or LrpC) also increased intra- and interspecies CT (nonself-DNA, ~8% nucleotide sequence divergence) in rec cells, but differentially reduced both types of CTs in certain rec-deficient strains. We describe that rok inactivation significantly stimulates intra and interspecies CT but differentially reduces them in transformation-deficient cells, perhaps by altering the nucleoid architecture. We extend the observation to other NAPs (Hbsu, LrpC).

摘要

Rok 是枯草芽孢杆菌核区相关蛋白(NAP),它负调控感受态发育并沉默异源基因。我们发现 rok 失活在不同程度上增加了 rpoB482 天然种内染色体转化(CT)和质粒转化。在 ΔaddAB、ΔrecO、recF15、ΔrecU、ΔruvAB 或 rec 细胞中,与 rok 细胞相比,种内 CT 显著增加,但 ΔrecD2 突变减少,而 ΔrecX、ΔradA 或 ΔdprA 突变则进一步降低了 Δrok 背景下的 CT。这些观察结果支持这样的观点,即 rok 失活通过改变受体 DNA 的拓扑结构,在 rec 缺陷菌株中不同程度地影响同源 DNA 的整合,并在较小程度上影响感受态亚群的大小。其他 NAP(Hbsu 或 LrpC)的缺失也增加了 rec 细胞中的种内和种间 CT(非自身 DNA,~8%核苷酸序列差异),但在某些 rec 缺陷菌株中,这两种 CT 都不同程度地减少。我们发现 rok 失活显著刺激了种内和种间 CT,但在转化缺陷细胞中却降低了它们,这可能是通过改变核区结构。我们将这一观察结果扩展到其他 NAP(Hbsu、LrpC)。

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