Nuclear Import Receptor Inhibits Phase Separation of FUS through Binding to Multiple Sites

Takuya Yoshizawa, Rustam Ali, Jenny Jiou, Ho Yee Joyce Fung, Kathleen A. Burke, Seung Joong Kim, Yuan Lin, William B. Peeples, Daniel Saltzberg, Michael Soniat, Jordan M. Baumhardt, Rudolf Oldenbourg, Andrej Sali, Nicolas L. Fawzi, Michael K. Rosen, Yuh Min Chook

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Liquid-liquid phase separation (LLPS) is believed to underlie formation of biomolecular condensates, cellular compartments that concentrate macromolecules without surrounding membranes. Physical mechanisms that control condensate formation/dissolution are poorly understood. The RNA-binding protein fused in sarcoma (FUS) undergoes LLPS in vitro and associates with condensates in cells. We show that the importin karyopherin-β2/transportin-1 inhibits LLPS of FUS. This activity depends on tight binding of karyopherin-β2 to the C-terminal proline-tyrosine nuclear localization signal (PY-NLS) of FUS. Nuclear magnetic resonance (NMR) analyses reveal weak interactions of karyopherin-β2 with sequence elements and structural domains distributed throughout the entirety of FUS. Biochemical analyses demonstrate that most of these same regions also contribute to LLPS of FUS. The data lead to a model where high-affinity binding of karyopherin-β2 to the FUS PY-NLS tethers the proteins together, allowing multiple, distributed weak intermolecular contacts to disrupt FUS self-association, blocking LLPS. Karyopherin-β2 may act analogously to control condensates in diverse cellular contexts. Distributed, energetically weak interactions between a karyopherin and the FUS nuclear localization signal disrupt FUS phase separation.

Original languageEnglish (US)
JournalCell
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Karyopherins
Cell Nucleus Active Transport
Cytoplasmic and Nuclear Receptors
Phase separation
Sarcoma
Liquids
Nuclear Localization Signals
Proline
Tyrosine
RNA-Binding Proteins
Macromolecules
Contacts (fluid mechanics)
Magnetic Resonance Spectroscopy
Dissolution
Nuclear magnetic resonance
Membranes

Keywords

  • amyotrophic lateral sclerosis
  • biomolecular condensate
  • FUS
  • intrinsically disordered protein
  • karyopherin-β2
  • liquid-liquid phase separation
  • low-complexity sequences
  • PY-NLS
  • RNA granule
  • transportin-1

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Nuclear Import Receptor Inhibits Phase Separation of FUS through Binding to Multiple Sites. / Yoshizawa, Takuya; Ali, Rustam; Jiou, Jenny; Fung, Ho Yee Joyce; Burke, Kathleen A.; Kim, Seung Joong; Lin, Yuan; Peeples, William B.; Saltzberg, Daniel; Soniat, Michael; Baumhardt, Jordan M.; Oldenbourg, Rudolf; Sali, Andrej; Fawzi, Nicolas L.; Rosen, Michael K.; Chook, Yuh Min.

In: Cell, 01.01.2018.

Research output: Contribution to journalArticle

Yoshizawa, T, Ali, R, Jiou, J, Fung, HYJ, Burke, KA, Kim, SJ, Lin, Y, Peeples, WB, Saltzberg, D, Soniat, M, Baumhardt, JM, Oldenbourg, R, Sali, A, Fawzi, NL, Rosen, MK & Chook, YM 2018, 'Nuclear Import Receptor Inhibits Phase Separation of FUS through Binding to Multiple Sites', Cell. https://doi.org/10.1016/j.cell.2018.03.003
Yoshizawa, Takuya ; Ali, Rustam ; Jiou, Jenny ; Fung, Ho Yee Joyce ; Burke, Kathleen A. ; Kim, Seung Joong ; Lin, Yuan ; Peeples, William B. ; Saltzberg, Daniel ; Soniat, Michael ; Baumhardt, Jordan M. ; Oldenbourg, Rudolf ; Sali, Andrej ; Fawzi, Nicolas L. ; Rosen, Michael K. ; Chook, Yuh Min. / Nuclear Import Receptor Inhibits Phase Separation of FUS through Binding to Multiple Sites. In: Cell. 2018.
@article{53b438bf211e4ff68a23b15490a2931c,
title = "Nuclear Import Receptor Inhibits Phase Separation of FUS through Binding to Multiple Sites",
abstract = "Liquid-liquid phase separation (LLPS) is believed to underlie formation of biomolecular condensates, cellular compartments that concentrate macromolecules without surrounding membranes. Physical mechanisms that control condensate formation/dissolution are poorly understood. The RNA-binding protein fused in sarcoma (FUS) undergoes LLPS in vitro and associates with condensates in cells. We show that the importin karyopherin-β2/transportin-1 inhibits LLPS of FUS. This activity depends on tight binding of karyopherin-β2 to the C-terminal proline-tyrosine nuclear localization signal (PY-NLS) of FUS. Nuclear magnetic resonance (NMR) analyses reveal weak interactions of karyopherin-β2 with sequence elements and structural domains distributed throughout the entirety of FUS. Biochemical analyses demonstrate that most of these same regions also contribute to LLPS of FUS. The data lead to a model where high-affinity binding of karyopherin-β2 to the FUS PY-NLS tethers the proteins together, allowing multiple, distributed weak intermolecular contacts to disrupt FUS self-association, blocking LLPS. Karyopherin-β2 may act analogously to control condensates in diverse cellular contexts. Distributed, energetically weak interactions between a karyopherin and the FUS nuclear localization signal disrupt FUS phase separation.",
keywords = "amyotrophic lateral sclerosis, biomolecular condensate, FUS, intrinsically disordered protein, karyopherin-β2, liquid-liquid phase separation, low-complexity sequences, PY-NLS, RNA granule, transportin-1",
author = "Takuya Yoshizawa and Rustam Ali and Jenny Jiou and Fung, {Ho Yee Joyce} and Burke, {Kathleen A.} and Kim, {Seung Joong} and Yuan Lin and Peeples, {William B.} and Daniel Saltzberg and Michael Soniat and Baumhardt, {Jordan M.} and Rudolf Oldenbourg and Andrej Sali and Fawzi, {Nicolas L.} and Rosen, {Michael K.} and Chook, {Yuh Min}",
year = "2018",
month = "1",
day = "1",
doi = "10.1016/j.cell.2018.03.003",
language = "English (US)",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",

}

TY - JOUR

T1 - Nuclear Import Receptor Inhibits Phase Separation of FUS through Binding to Multiple Sites

AU - Yoshizawa, Takuya

AU - Ali, Rustam

AU - Jiou, Jenny

AU - Fung, Ho Yee Joyce

AU - Burke, Kathleen A.

AU - Kim, Seung Joong

AU - Lin, Yuan

AU - Peeples, William B.

AU - Saltzberg, Daniel

AU - Soniat, Michael

AU - Baumhardt, Jordan M.

AU - Oldenbourg, Rudolf

AU - Sali, Andrej

AU - Fawzi, Nicolas L.

AU - Rosen, Michael K.

AU - Chook, Yuh Min

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Liquid-liquid phase separation (LLPS) is believed to underlie formation of biomolecular condensates, cellular compartments that concentrate macromolecules without surrounding membranes. Physical mechanisms that control condensate formation/dissolution are poorly understood. The RNA-binding protein fused in sarcoma (FUS) undergoes LLPS in vitro and associates with condensates in cells. We show that the importin karyopherin-β2/transportin-1 inhibits LLPS of FUS. This activity depends on tight binding of karyopherin-β2 to the C-terminal proline-tyrosine nuclear localization signal (PY-NLS) of FUS. Nuclear magnetic resonance (NMR) analyses reveal weak interactions of karyopherin-β2 with sequence elements and structural domains distributed throughout the entirety of FUS. Biochemical analyses demonstrate that most of these same regions also contribute to LLPS of FUS. The data lead to a model where high-affinity binding of karyopherin-β2 to the FUS PY-NLS tethers the proteins together, allowing multiple, distributed weak intermolecular contacts to disrupt FUS self-association, blocking LLPS. Karyopherin-β2 may act analogously to control condensates in diverse cellular contexts. Distributed, energetically weak interactions between a karyopherin and the FUS nuclear localization signal disrupt FUS phase separation.

AB - Liquid-liquid phase separation (LLPS) is believed to underlie formation of biomolecular condensates, cellular compartments that concentrate macromolecules without surrounding membranes. Physical mechanisms that control condensate formation/dissolution are poorly understood. The RNA-binding protein fused in sarcoma (FUS) undergoes LLPS in vitro and associates with condensates in cells. We show that the importin karyopherin-β2/transportin-1 inhibits LLPS of FUS. This activity depends on tight binding of karyopherin-β2 to the C-terminal proline-tyrosine nuclear localization signal (PY-NLS) of FUS. Nuclear magnetic resonance (NMR) analyses reveal weak interactions of karyopherin-β2 with sequence elements and structural domains distributed throughout the entirety of FUS. Biochemical analyses demonstrate that most of these same regions also contribute to LLPS of FUS. The data lead to a model where high-affinity binding of karyopherin-β2 to the FUS PY-NLS tethers the proteins together, allowing multiple, distributed weak intermolecular contacts to disrupt FUS self-association, blocking LLPS. Karyopherin-β2 may act analogously to control condensates in diverse cellular contexts. Distributed, energetically weak interactions between a karyopherin and the FUS nuclear localization signal disrupt FUS phase separation.

KW - amyotrophic lateral sclerosis

KW - biomolecular condensate

KW - FUS

KW - intrinsically disordered protein

KW - karyopherin-β2

KW - liquid-liquid phase separation

KW - low-complexity sequences

KW - PY-NLS

KW - RNA granule

KW - transportin-1

UR - http://www.scopus.com/inward/record.url?scp=85044718274&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85044718274&partnerID=8YFLogxK

U2 - 10.1016/j.cell.2018.03.003

DO - 10.1016/j.cell.2018.03.003

M3 - Article

C2 - 29677513

AN - SCOPUS:85044718274

JO - Cell

JF - Cell

SN - 0092-8674

ER -