Probing DNA-Amyloid Interaction and Gel Formation by Active Magnetic Wire Microrheology

Methods Mol Biol. 2022:2538:285-303. doi: 10.1007/978-1-0716-2529-3_19.

Abstract

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.

Keywords: Bacterial amyloid; Magnetic rotational spectroscopy; Magnetic wire; Microrheology; Nucleoid-associated protein.

MeSH terms

  • Amyloid* / chemistry
  • Amyloidogenic Proteins*
  • Bacterial Proteins / metabolism
  • DNA
  • DNA Probes
  • Magnetic Phenomena
  • Viscosity

Substances

  • Amyloid
  • Amyloidogenic Proteins
  • Bacterial Proteins
  • DNA Probes
  • DNA