SUMOylation of the brain-predominant Ataxin-3 isoform modulates its interaction with p97

Almeida, Bruno; Abreu, Isabel A.; Matos, Carlos A.; Fraga, Joana S.; Fernandes, Sara; Macedo, Maria G.; Gutiérrez-Gallego, Ricardo; Pereira, Pedro José Barbosa; Carvalho, Ana Luı́sa; Macedo-Ribeiro, Sandra

doi:10.1016/j.bbadis.2015.06.010

Cited by 34 publications

(29 citation statements)

References 66 publications

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“…Our affinity measurements are also consistent with ITC studies that measured the binding of the ataxin3 VBM peptide to the p97 N-domain, reporting a K D of 15.6 M (84). In contrast, our measurements are not consistent with a previous report in which SPR was used to derive a substantially higher affinity estimate for the p97-ataxin3 interaction (K D ϳ1 nM) (85). However, as we have shown in this work, the p97-ataxin3 system presents challenges when using SPR to measure binding affinities, and if mass transport and rebinding events are not accounted for appropriately, the affinity and binding kinetics will not be calculated correctly (see supplemental data, SPR experimental details, and supplemental Figs.…”

Section: Discussioncontrasting

confidence: 99%

“…The experiments described in Ref. 85 feature high ligand immobilization density, low analyte flow rates, and short dissociation times, all conditions likely to promote surface-induced artifacts, which are reflected in poor global fits to the kinetic data. Hence, the apparent disagreement between our affinity measurements and those reported in the earlier paper (85) may simply reflect differences in data analysis and is not a genuine difference in the behavior of the proteins.…”

Section: Discussionmentioning

confidence: 99%

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Interaction between the AAA+ ATPase p97 and its cofactor ataxin3 in health and disease: Nucleotide-induced conformational changes regulate cofactor binding

Rao

Williams

Simon

et al. 2017

Journal of Biological Chemistry

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p97 is an essential ATPase associated with various cellular activities (AAA) that functions as a segregase in diverse cellular processes, including the maintenance of proteostasis. p97 interacts with different cofactors that target it to distinct pathways; an important example is the deubiquitinase ataxin3, which collaborates with p97 in endoplasmic reticulum-associated degradation. However, the molecular details of this interaction have been unclear. Here, we characterized the binding of ataxin3 to p97, showing that ataxin3 binds with low-micromolar affinity to both wild-type p97 and mutants linked to degenerative disorders known as multisystem proteinopathy 1 (MSP1); we further showed that the stoichiometry of binding is one ataxin3 molecule per p97 hexamer. We mapped the binding determinants on each protein, demonstrating that ataxin3's p97/VCP-binding motif interacts with the inter-lobe cleft in the N-domain of p97. We also probed the nucleotide dependence of this interaction, confirming that ataxin3 and p97 associate in the presence of ATP and in the absence of nucleotide, but not in the presence of ADP. Our experiments suggest that an ADP-driven downward movement of the p97 N-terminal domain dislodges ataxin3 by inducing a steric clash between the D1-domain and ataxin3's C terminus. In contrast, MSP1 mutants of p97 bind ataxin3 irrespective of their nucleotide state, indicating a failure by these mutants to translate ADP binding into a movement of the N-terminal domain. Our model provides a mechanistic explanation for how nucleotides regulate the p97-ataxin3 interaction and why atypical cofactor binding is observed with MSP1 mutants.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Interaction between the AAA+ ATPase p97 and its cofactor ataxin3 in health and disease: Nucleotide-induced conformational changes regulate cofactor binding

Rao

Williams

Simon

et al. 2017

Journal of Biological Chemistry

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“…Endogenous VCP co-localizes with the polyglutamine-containing aggregates in patients with HD and Machado–Joseph disease232425. VCP can bind directly to multiple polyglutamine disease proteins, including huntingtin, ataxin-1, ataxin-7 and androgen receptors2627.…”

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confidence: 99%

VCP recruitment to mitochondria causes mitophagy impairment and neurodegeneration in models of Huntington’s disease

et al. 2016

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Mutant Huntingtin (mtHtt) causes neurodegeneration in Huntington's disease (HD) by evoking defects in the mitochondria, but the underlying mechanisms remains elusive. Our proteomic analysis identifies valosin-containing protein (VCP) as an mtHtt-binding protein on the mitochondria. Here we show that VCP is selectively translocated to the mitochondria, where it is bound to mtHtt in various HD models. Mitochondria-accumulated VCP elicits excessive mitophagy, causing neuronal cell death. Blocking mtHtt/VCP mitochondrial interaction with a peptide, HV-3, abolishes VCP translocation to the mitochondria, corrects excessive mitophagy and reduces cell death in HD mouse- and patient-derived cells and HD transgenic mouse brains. Treatment with HV-3 reduces behavioural and neuropathological phenotypes of HD in both fragment- and full-length mtHtt transgenic mice. Our findings demonstrate a causal role of mtHtt-induced VCP mitochondrial accumulation in HD pathogenesis and suggest that the peptide HV-3 might be a useful tool for developing new therapeutics to treat HD.

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“…Mechanisms that modulate the lifespan and stabilization of p97-cofactor complexes like PTMs of p97 (Ewens et al, 2010) and its cofactors (Uchiyama et al, 2003; Almeida et al, 2015) or substrate recruitment by cofactors have been proposed.…”

Section: Regulation Of P97—cofactor Assemblymentioning

confidence: 99%

“…Beside numerous phosphorylation, ubiquitylation and acetylation events also SUMOylation, palmitoylation, methylation, succinylation, and S-glutathionylation were identified (Figure 7A). In the case of p97 PTMs may promote or prevent protein–protein interactions by obscuring existing binding sites, generating new interfaces, or triggering conformational changes, with the exact functional consequence dictated by the substrate and cellular context (Almeida et al, 2015). …”

Section: Regulation Of P97—cofactor Assemblymentioning

confidence: 99%

The Interplay of Cofactor Interactions and Post-translational Modifications in the Regulation of the AAA+ ATPase p97

Hänzelmann

Schindelin

2017

Front. Mol. Biosci.

129

134

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The hexameric type II AAA ATPase (ATPase associated with various activities) p97 (also referred to as VCP, Cdc48, and Ter94) is critically involved in a variety of cellular activities including pathways such as DNA replication and repair which both involve chromatin remodeling, and is a key player in various protein quality control pathways mediated by the ubiquitin proteasome system as well as autophagy. Correspondingly, p97 has been linked to various pathophysiological states including cancer, neurodegeneration, and premature aging. p97 encompasses an N-terminal domain, two highly conserved ATPase domains and an unstructured C-terminal tail. This enzyme hydrolyzes ATP and utilizes the resulting energy to extract or disassemble protein targets modified with ubiquitin from stable protein assemblies, chromatin and membranes. p97 participates in highly diverse cellular processes and hence its activity is tightly controlled. This is achieved by multiple regulatory cofactors, which either associate with the N-terminal domain or interact with the extreme C-terminus via distinct binding elements and target p97 to specific cellular pathways, sometimes requiring the simultaneous association with more than one cofactor. Most cofactors are recruited to p97 through conserved binding motifs/domains and assist in substrate recognition or processing by providing additional molecular properties. A tight control of p97 cofactor specificity and diversity as well as the assembly of higher-order p97-cofactor complexes is accomplished by various regulatory mechanisms, which include bipartite binding, binding site competition, changes in oligomeric assemblies, and nucleotide-induced conformational changes. Furthermore, post-translational modifications (PTMs) like acetylation, palmitoylation, phosphorylation, SUMOylation, and ubiquitylation of p97 have been reported which further modulate its diverse molecular activities. In this review, we will describe the molecular basis of p97-cofactor specificity/diversity and will discuss how PTMs can modulate p97-cofactor interactions and affect the physiological and patho-physiological functions of p97.

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SUMOylation of the brain-predominant Ataxin-3 isoform modulates its interaction with p97

Abstract: These findings highlight the role of SUMOylation as a regulator of Atx3 function, with implications on Atx3 protein interaction network and self-assembly, with potential impact for further understanding the molecular mechanisms underlying MJD pathogenesis.

Cited by 34 publications

References 66 publications

Interaction between the AAA+ ATPase p97 and its cofactor ataxin3 in health and disease: Nucleotide-induced conformational changes regulate cofactor binding

Interaction between the AAA+ ATPase p97 and its cofactor ataxin3 in health and disease: Nucleotide-induced conformational changes regulate cofactor binding

VCP recruitment to mitochondria causes mitophagy impairment and neurodegeneration in models of Huntington’s disease

The Interplay of Cofactor Interactions and Post-translational Modifications in the Regulation of the AAA+ ATPase p97

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