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Phase Separation of FUS with Poly(ADP-ribosyl)ated PARP1 Is Controlled by Polyamines, Divalent Metal Cations, and Poly(ADP-ribose) Structure
by
Hamon, Loic
, Singatulina, Anastasia S.
, Sukhanova, Maria V.
, Lavrik, Olga I.
, Anarbaev, Rashid O.
, Naumenko, Konstantin N.
, Pastré, David
in
Cations, Divalent
/ DNA damage
/ DNA repair
/ Enzymes
/ Humans
/ Metabolism
/ Microscopy
/ Microscopy, Atomic Force
/ Phase Separation
/ Poly (ADP-Ribose) Polymerase-1 - chemistry
/ Poly (ADP-Ribose) Polymerase-1 - metabolism
/ Poly Adenosine Diphosphate Ribose - chemistry
/ Poly Adenosine Diphosphate Ribose - metabolism
/ Poly ADP Ribosylation
/ Polyamines
/ Polyamines - chemistry
/ Polyamines - metabolism
/ Proteins
/ RNA-Binding Protein FUS - chemistry
/ RNA-Binding Protein FUS - genetics
/ RNA-Binding Protein FUS - metabolism
/ Sarcoma
2024
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Phase Separation of FUS with Poly(ADP-ribosyl)ated PARP1 Is Controlled by Polyamines, Divalent Metal Cations, and Poly(ADP-ribose) Structure
by
Hamon, Loic
, Singatulina, Anastasia S.
, Sukhanova, Maria V.
, Lavrik, Olga I.
, Anarbaev, Rashid O.
, Naumenko, Konstantin N.
, Pastré, David
in
Cations, Divalent
/ DNA damage
/ DNA repair
/ Enzymes
/ Humans
/ Metabolism
/ Microscopy
/ Microscopy, Atomic Force
/ Phase Separation
/ Poly (ADP-Ribose) Polymerase-1 - chemistry
/ Poly (ADP-Ribose) Polymerase-1 - metabolism
/ Poly Adenosine Diphosphate Ribose - chemistry
/ Poly Adenosine Diphosphate Ribose - metabolism
/ Poly ADP Ribosylation
/ Polyamines
/ Polyamines - chemistry
/ Polyamines - metabolism
/ Proteins
/ RNA-Binding Protein FUS - chemistry
/ RNA-Binding Protein FUS - genetics
/ RNA-Binding Protein FUS - metabolism
/ Sarcoma
2024
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Phase Separation of FUS with Poly(ADP-ribosyl)ated PARP1 Is Controlled by Polyamines, Divalent Metal Cations, and Poly(ADP-ribose) Structure
by
Hamon, Loic
, Singatulina, Anastasia S.
, Sukhanova, Maria V.
, Lavrik, Olga I.
, Anarbaev, Rashid O.
, Naumenko, Konstantin N.
, Pastré, David
in
Cations, Divalent
/ DNA damage
/ DNA repair
/ Enzymes
/ Humans
/ Metabolism
/ Microscopy
/ Microscopy, Atomic Force
/ Phase Separation
/ Poly (ADP-Ribose) Polymerase-1 - chemistry
/ Poly (ADP-Ribose) Polymerase-1 - metabolism
/ Poly Adenosine Diphosphate Ribose - chemistry
/ Poly Adenosine Diphosphate Ribose - metabolism
/ Poly ADP Ribosylation
/ Polyamines
/ Polyamines - chemistry
/ Polyamines - metabolism
/ Proteins
/ RNA-Binding Protein FUS - chemistry
/ RNA-Binding Protein FUS - genetics
/ RNA-Binding Protein FUS - metabolism
/ Sarcoma
2024
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Phase Separation of FUS with Poly(ADP-ribosyl)ated PARP1 Is Controlled by Polyamines, Divalent Metal Cations, and Poly(ADP-ribose) Structure
Journal Article
Phase Separation of FUS with Poly(ADP-ribosyl)ated PARP1 Is Controlled by Polyamines, Divalent Metal Cations, and Poly(ADP-ribose) Structure
2024
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Overview
Fused in sarcoma (FUS) is involved in the formation of nuclear biomolecular condensates associated with poly(ADP-ribose) [PAR] synthesis catalyzed by a DNA damage sensor such as PARP1. Here, we studied FUS microphase separation induced by poly(ADP-ribosyl)ated PARP1WT [PAR-PARP1WT] or its catalytic variants PARP1Y986S and PARP1Y986H, respectively, synthesizing (short PAR)-PARP1Y986S or (short hyperbranched PAR)-PARP1Y986H using dynamic light scattering, fluorescence microscopy, turbidity assays, and atomic force microscopy. We observed that biologically relevant cations such as Mg2+, Ca2+, or Mn2+ or polyamines (spermine4+ or spermidine3+) were essential for the assembly of FUS with PAR-PARP1WT and FUS with PAR-PARP1Y986S in vitro. We estimated the range of the FUS-to-PAR-PARP1 molar ratio and the cation concentration that are favorable for the stability of the protein’s microphase-separated state. We also found that FUS microphase separation induced by PAR-PARP1Y986H (i.e., a PARP1 variant attaching short hyperbranched PAR to itself) can occur in the absence of cations. The dependence of PAR-PARP1-induced FUS microphase separation on cations and on the branching of the PAR structure points to a potential role of the latter in the regulation of the formation of FUS-related biological condensates and requires further investigation.
Publisher
MDPI AG,MDPI
Subject
/ Enzymes
/ Humans
/ Poly (ADP-Ribose) Polymerase-1 - chemistry
/ Poly (ADP-Ribose) Polymerase-1 - metabolism
/ Poly Adenosine Diphosphate Ribose - chemistry
/ Poly Adenosine Diphosphate Ribose - metabolism
/ Proteins
/ RNA-Binding Protein FUS - chemistry
/ RNA-Binding Protein FUS - genetics
/ RNA-Binding Protein FUS - metabolism
/ Sarcoma
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