Interplay of folded domains and the disordered low-complexity domain in mediating hnRNPA1 phase separation. Issue 5 (12th February 2021)

• Record Type: Journal Article
• Authors: Martin, Erik W; Thomasen, F Emil; Milkovic, Nicole M; Cuneo, Matthew J; Grace, Christy R; Nourse, Amanda; Lindorff-Larsen, Kresten; Mittag, Tanja
• Is Part Of: Nucleic acids research. Volume 49:Issue 5(2021)
• Page Start: 2931
• Page End: 2945
• Publication Date: 2021-02-12
• Subjects: Nucleic acids -- Periodicals; Molecular biology -- Periodicals; 572.805
• DOI: 10.1093/nar/gkab063
• Languages: English
• ISSNs: 0305-1048

Abstract

Abstract: Liquid–liquid phase separation underlies the membrane-less compartmentalization of cells. Intrinsically disordered low-complexity domains (LCDs) often mediate phase separation, but how their phase behavior is modulated by folded domains is incompletely understood. Here, we interrogate the interplay between folded and disordered domains of the RNA-binding protein hnRNPA1. The LCD of hnRNPA1 is sufficient for mediating phase separation in vitro . However, we show that the folded RRM domains and a folded solubility-tag modify the phase behavior, even in the absence of RNA. Notably, the presence of the folded domains reverses the salt dependence of the driving force for phase separation relative to the LCD alone. Small-angle X-ray scattering experiments and coarse-grained MD simulations show that the LCD interacts transiently with the RRMs and/or the solubility-tag in a salt-sensitive manner, providing a mechanistic explanation for the observed salt-dependent phase separation. These data point to two effects from the folded domains: (i) electrostatically-mediated interactions that compact hnRNPA1 and contribute to phase separation and (ii) increased solubility at higher ionic strengths mediated by the folded domains. The interplay between disordered and folded domains can modify the dependence of phase behavior on solution conditions and can obscure signatures of physicochemical interactions underlying phase separation. Graphical Abstract: