Interplay of folded domains and the disordered low-complexity domain in mediating hnRNPA1 phase separation
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Interplay of folded domains and the disordered low-complexity domain in mediating hnRNPA1 phase separation. / Martin, Erik W.; Thomasen, F. Emil; Milkovic, Nicole M.; Cuneo, Matthew J.; Grace, Christy R.; Nourse, Amanda; Lindorff-Larsen, Kresten; Mittag, Tanja.
In: Nucleic Acids Research, Vol. 49, No. 5, 2021, p. 2931-2945.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Interplay of folded domains and the disordered low-complexity domain in mediating hnRNPA1 phase separation
AU - Martin, Erik W.
AU - Thomasen, F. Emil
AU - Milkovic, Nicole M.
AU - Cuneo, Matthew J.
AU - Grace, Christy R.
AU - Nourse, Amanda
AU - Lindorff-Larsen, Kresten
AU - Mittag, Tanja
PY - 2021
Y1 - 2021
N2 - 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.
AB - 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.
U2 - 10.1093/nar/gkab063
DO - 10.1093/nar/gkab063
M3 - Journal article
C2 - 33577679
AN - SCOPUS:85103228607
VL - 49
SP - 2931
EP - 2945
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 5
ER -
ID: 260743480