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Several One-Domain Zinc Finger µ-Proteins of Haloferax Volcanii Are Important for Stress Adaptation Biofilm Formation and Swarming

机译:Haloferax Volcanii的几种一域锌指µ蛋白对于应力适应生物膜形成和形成群很重要

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摘要

Zinc finger domains are highly structured and can mediate interactions to DNA, RNA, proteins, lipids, and small molecules. Accordingly, zinc finger proteins are very versatile and involved in many biological functions. Eukaryotes contain a wealth of zinc finger proteins, but zinc finger proteins have also been found in archaea and bacteria. Large zinc finger proteins have been well studied, however, in stark contrast, single domain zinc finger µ-proteins of less than 70 amino acids have not been studied at all, with one single exception. Therefore, 16 zinc finger µ-proteins of the haloarchaeon Haloferax volcanii were chosen and in frame deletion mutants of the cognate genes were generated. The phenotypes of mutants and wild-type were compared under eight different conditions, which were chosen to represent various pathways and involve many genes. None of the mutants differed from the wild-type under optimal or near-optimal conditions. However, 12 of the 16 mutants exhibited a phenotypic difference under at least one of the four following conditions: Growth in synthetic medium with glycerol, growth in the presence of bile acids, biofilm formation, and swarming. In total, 16 loss of function and 11 gain of function phenotypes were observed. Five mutants indicated counter-regulation of a sessile versus a motile life style in H. volcanii. In conclusion, the generation and analysis of a set of deletion mutants demonstrated the high importance of zinc finger µ-proteins for various biological functions, and it will be the basis for future mechanistic insight.
机译:锌指结构域高度结构化,可以介导与DNA,RNA,蛋白质,脂质和小分子的相互作用。因此,锌指蛋白非常通用并且涉及许多生物学功能。真核生物含有大量的锌指蛋白,但是锌指蛋白也存在于古细菌和细菌中。大型锌指蛋白已被很好地研究,但是,与之形成鲜明对比的是,除了一个单一例外,根本没有研究少于70个氨基酸的单结构域锌指µ蛋白。因此,选择了卤代古细菌Haloferax volcanii的16个锌指μ蛋白,并在同源缺失基因的框架内缺失突变体。在八个不同条件下比较了突变体的表型和野生型,选择它们代表各种途径并涉及许多基因。在最佳或接近最佳条件下,所有突变体均与野生型没有区别。但是,在以下四个条件中的至少一个条件下,这16个突变体中的12个表现出表型差异:在含甘油的合成培养基中生长,在胆汁酸存在下生长,生物膜形成和成群。总共观察到16种功能丧失和11种功能表现型。五个突变体表明在火山嗜血杆菌中无柄相对于运动型生活方式的反调节。总之,一组缺失突变体的产生和分析证明了锌指µ蛋白对于各种生物学功能的高度重要性,它将为将来的机理研究提供基础。

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