Single molecule probing of disordered RNA binding proteins

Kevin Rhine; Sua Myong

doi:10.1016/j.xpro.2022.101131

Single molecule probing of disordered RNA binding proteins

STAR Protoc. 2022 Jan 28;3(1):101131. doi: 10.1016/j.xpro.2022.101131. eCollection 2022 Mar 18.

Authors

Kevin Rhine^{1

2}, Sua Myong^{1

3

4}

Affiliations

¹ Program in Cell, Molecular, Developmental Biology and Biophysics, Johns Hopkins University, Baltimore, MD 21218, USA.
² Department of Biology, Johns Hopkins University, Baltimore, MD 21218, USA.
³ Department of Biophysics, Johns Hopkins University, Baltimore, MD 21218, USA.
⁴ Physics Frontier Center (Center for the Physics of Living Cells), University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.

Abstract

Liquid-liquid phase separation of intrinsically disordered proteins is known to underlie diverse pathologies such as neurodegeneration, cancer, and aging. The nucleation step of condensate formation is of critical importance for understanding how healthy and disease-associated condensates differ. Here, we describe four orthogonal single-molecule techniques that enable molecular tracking of the RNA-protein interaction, RNA-induced oligomerization, and kinetics of nucleation. These approaches allow researchers to directly interrogate the initial steps of liquid-liquid phase separation. For complete details on the use and execution of this profile, please refer to Niaki et al. (2020), Rhine et al. (2020), and Rhine et al. (2022).

Keywords: Biophysics; Biotechnology and bioengineering; Molecular Biology; Single-molecule Assays.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Biophysical Phenomena
Intrinsically Disordered Proteins* / chemistry
Kinetics
RNA
RNA-Binding Proteins* / chemistry

Substances

Intrinsically Disordered Proteins
RNA-Binding Proteins
RNA

Grants and funding

F31 NS113439/NS/NINDS NIH HHS/United States