Radiom Milad, Oikonomou Evdokia K, Grados Arnaud, Receveur Mathieu, Berret Jean-François
Université de Paris, CNRS, Paris, France.
Institute for Food, Nutrition and Health, D-HEST, ETH Zürich, Zürich, Switzerland.
Methods Mol Biol. 2022;2538:285-303. doi: 10.1007/978-1-0716-2529-3_19.
Recent studies have shown that bacterial nucleoid-associated proteins (NAPs) can bind to DNA and result in altered structural organization and bridging interactions. Under spontaneous self-assembly, NAPs may also form anisotropic amyloid fibers, whose effects are still more significant on DNA dynamics. To test this hypothesis, microrheology experiments on dispersions of DNA associated with the amyloid terminal domain (CTR) of the bacterial protein Hfq were performed using magnetic rotational spectroscopy (MRS). In this chapter, we survey this microrheology technique based on the remote actuation of magnetic wires embedded in a sample. MRS is interesting as it is easy to implement and does not require complex procedures regarding data treatment. Pertaining to the interaction between DNA and amyloid fibers, it is found that DNA and Hfq-CTR protein dispersions behave like a gel, an outcome that suggests the formation of a network of amyloid fibers cross-linked with the DNA strands. In contrast, the pristine DNA and Hfq-CTR dispersions behave as purely viscous liquids. To broaden the scope of the MRS technique, we include theoretical predictions for the rotation of magnetic wires regarding the generic behaviors of basic rheological models from continuum mechanics, and we list the complex fluids studied by this technique over the past 10 years.
最近的研究表明,细菌类核相关蛋白(NAPs)可与DNA结合,导致结构组织改变和桥连相互作用。在自发自组装过程中,NAPs还可能形成各向异性的淀粉样纤维,其对DNA动力学的影响更为显著。为验证这一假设,利用磁旋转光谱(MRS)对与细菌蛋白Hfq的淀粉样末端结构域(CTR)相关的DNA分散体进行了微观流变学实验。在本章中,我们将基于对嵌入样品中的磁丝的远程驱动来概述这种微观流变学技术。MRS很有意思,因为它易于实施,且在数据处理方面不需要复杂的程序。关于DNA与淀粉样纤维之间的相互作用,发现DNA和Hfq-CTR蛋白分散体表现得像凝胶,这一结果表明形成了与DNA链交联的淀粉样纤维网络。相比之下,原始的DNA和Hfq-CTR分散体表现为纯粘性液体。为拓宽MRS技术的应用范围,我们纳入了关于磁丝旋转的理论预测,这些预测涉及连续介质力学中基本流变模型的一般行为,并且我们列出了过去10年中用该技术研究的复杂流体。