Assembly Dynamics and Stoichiometry of the Apoptosis Signal-regulating Kinase (ASK) Signalosome in Response to Electrophile Stress

Federspiel, Joel D.; Codreanu, Simona G.; Palubinsky, Amy M.; Winland, Ama J.; Betanzos, Carlos Morales; McLaughlin, BethAnn; Liebler, D.C.

doi:10.1074/mcp.m115.057364

Cited by 33 publications

(35 citation statements)

References 69 publications

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“…This behaviour suggests that the SAM domains of each protein contribute markedly to the heterotypic ASK1-ASK2 complexes previously reported (4)(5)(6)(7)(8)28).…”

Section: Figure 3: Heterotypic Interactions Between Ask Sam Domains (supporting

confidence: 69%

“…When transfected together, wild-type ASK1 and ASK2 show significantly lower basal levels of kinase activity, which is nonetheless activated by HNE. Based on the complete lack of signal in cells transfected with ASK2 alone (WT or C1268E; Figure 4A/B central lanes), the phospho-ASK signal is specific to ASK1, rather than cross-reactivity of the ASK1- Elegant endogenous mass spectrometry studies have previously shown that ASK1 associates with near-stoichiometric amounts of ASK2 (28). We employed an orthogonal approach-bimolecular complementation affinity purification (BiCAP (29,30))-to determine if stoichiometric association of ASK1-ASK2 occurs as part of larger hetero-oligomeric complex.…”

Section: Sam-mediated Oligomers Regulate Activity and Stoichiometry Imentioning

confidence: 98%

“…ASK1 and ASK2 have previously been reported to associate through their C-terminal domains (4), and endogenous ASK1 and ASK2 have been reported to exist in complex with one another at an equal ratio (28). We next sought to test whether such heterotypic association could be driven through the isolated SAM domains of each ASK protein.…”

Section: Ask1-ask2 Form Heterotypic Complexes Via the Ml-eh Interfacementioning

confidence: 99%

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Mechanism of preferential complex formation by Apoptosis Signal-regulating Kinases

Burgess

et al. 2019

Preprint

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Apoptosis signal-regulating kinases (ASK1-3) are activators of the P38 and JNK MAP kinase pathways. ASK1-3 form oligomeric complexes known as ASK signalosomes that initiate signalling cascades in response to diverse stress stimuli. Here we demonstrate that oligomerization of ASK proteins is driven by previously uncharacterised sterile-alpha motif (SAM) domains that reside at the C-terminus of each ASK protein. SAM domains from ASK1-3 have distinct behaviours: ASK1 forms unstable oligomers, ASK2 is predominantly monomeric, and the ASK3 SAM domain forms a stable oligomer even at low concentration.In contrast to their isolated behaviour, the ASK1 and ASK2 SAM domains preferentially form a stable heterocomplex. The crystal structure of the ASK3 SAM domain, small-angle X-ray scattering, and mutagenesis suggests that ASK3 oligomers and ASK1-ASK2 complexes form discrete quasi-helical rings, via the mid-loop-end-helix interface. Preferential ASK1-ASK2 binding is consistent with mass spectrometry showing that full-length ASK1 forms heterooligomeric complexes incorporating high levels of ASK2. Accordingly, disruption of SAM domain-association impairs ASK activity in the context of electrophilic stress induced by 4-hydroxy-2-nonenal. These findings provide a structural template for how ASK proteins assemble foci to drive inflammatory signalling, and reinforce that strategies targeting ASK kinases should consider the concerted actions of multiple ASK family members.

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“…This behaviour suggests that the SAM domains of each protein contribute markedly to the heterotypic ASK1-ASK2 complexes previously reported (4)(5)(6)(7)(8)28).…”

Section: Figure 3: Heterotypic Interactions Between Ask Sam Domains (supporting

confidence: 69%

Section: Sam-mediated Oligomers Regulate Activity and Stoichiometry Imentioning

confidence: 98%

Section: Ask1-ask2 Form Heterotypic Complexes Via the Ml-eh Interfacementioning

confidence: 99%

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Mechanism of preferential complex formation by Apoptosis Signal-regulating Kinases

Burgess

et al. 2019

Preprint

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“…SAINT assigns each protein a SAINT Probability Score ("P-Score"), on a scale of 0 to 1, with 1 being the top score. Based upon review of the literature, we chose 0.65 as a SAINT cut-off score for enrichment (68)(69)(70)(71)(72)(73). With the approach used here, all proteins have two P-Scores, one each for basal or insulin enrichment over the respective negative controls.…”

Section: Resultsmentioning

confidence: 99%

Characterization of the CLASP2 Protein Interaction Network Identifies SOGA1 as a Microtubule-Associated Protein

Kruse

Krantz

Barker

et al. 2017

Molecular & Cellular Proteomics

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SummaryCLASP2 is a microtubule-associated protein that undergoes insulin-stimulated phosphorylation and co-localization with reorganized actin and GLUT4 at the plasma membrane.To gain insight to the role of CLASP2 in this system, we developed and successfully executed a streamlined interactome approach and built a CLASP2 protein network in 3T3-L1 adipocytes.Using two different commercially available antibodies for CLASP2 and an antibody for epitopetagged, overexpressed CLASP2, we performed multiple affinity purification coupled with mass spectrometry (AP-MS) experiments in combination with label-free quantitative proteomics and analyzed the data with the bioinformatics tool Significance Analysis of Interactome (SAINT). We discovered that CLASP2 co-immunoprecipitates (co-IPs) the novel protein SOGA1, the microtubule-associated protein kinase MARK2, and the microtubule/actin-regulating protein G2L1. The GTPase-activating proteins AGAP1 and AGAP3 were also enriched in the CLASP2 interactome, although subsequent AGAP3 and CLIP2 interactome analysis suggests a preference of AGAP3 for CLIP2. Follow-up MARK2 interactome analysis confirmed reciprocal co-IP of CLASP2 and also revealed MARK2 can co-IP SOGA1, glycogen synthase, and glycogenin.Investigating the SOGA1 interactome confirmed SOGA1 can reciprocal co-IP both CLASP2 and MARK2 as well as glycogen synthase and glycogenin. SOGA1 was confirmed to colocalize with CLASP2 and also with tubulin, which identifies SOGA1 as a new microtubule-associated protein.These results introduce the metabolic function of these proposed novel protein networks and their relationship with microtubules as new fields of cytoskeleton-associated protein biology. 5

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“…1A). The active signaling form of ASK1 in cells is thought to be an oligomer often referred to as the "ASK signalosome," which also contains ASK2 and ASK3 (20)(21)(22). To the N terminus of the kinase domain lie several regions with regulatory roles but poorly understood structures.…”

mentioning

confidence: 99%

Structural basis of autoregulatory scaffolding by apoptosis signal-regulating kinase 1

Weijman

Kumar

Jamieson

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Apoptosis signal-regulating kinases (ASK1-3) are apical kinases of the p38 and JNK MAP kinase pathways. They are activated by diverse stress stimuli, including reactive oxygen species, cytokines, and osmotic stress; however, a molecular understanding of how ASK proteins are controlled remains obscure. Here, we report a biochemical analysis of the ASK1 kinase domain in conjunction with its N-terminal thioredoxin-binding domain, along with a central regulatory region that links the two. We show that in solution the central regulatory region mediates a compact arrangement of the kinase and thioredoxin-binding domains and the central regulatory region actively primes MKK6, a key ASK1 substrate, for phosphorylation. The crystal structure of the central regulatory region reveals an unusually compact tetratricopeptide repeat (TPR) region capped by a cryptic pleckstrin homology domain. Biochemical assays show that both a conserved surface on the pleckstrin homology domain and an intact TPR region are required for ASK1 activity. We propose a model in which the central regulatory region promotes ASK1 activity via its pleckstrin homology domain but also facilitates ASK1 autoinhibition by bringing the thioredoxin-binding and kinase domains into close proximity. Such an architecture provides a mechanism for control of ASK-type kinases by diverse activators and inhibitors and demonstrates an unexpected level of autoregulatory scaffolding in mammalian stress-activated MAP kinase signaling.itogen-activated protein (MAP) kinase cascades transmit signals from membrane-associated receptors to intracellular targets to effect changes in cellular behavior. They form a hierarchical system in which activated upstream kinases (MAP3Ks) phosphorylate intermediate MAP kinase kinases (MAP2Ks), which in turn phosphorylate terminal MAP kinases, primarily ERK, p38, and JNK and their isoforms (1). Extensive studies have focused on the activation of RAS-RAF-MEK upstream in the ERK pathway and provided fertile ground for the discovery of new therapeutics (2). In contrast to the ERK pathway, which primarily promotes cellular proliferation, JNK and p38 phosphorylate a range of substrates to promote inflammation and cell death (1, 3). In addition, cross-regulation among the p38, JNK, and ERK pathways is important for the efficacy of various cancer therapies that are in use or in development (4, 5). Molecular details on the more diverse upstream regulation of the p38 and JNK pathways are currently less clear, however.Apoptosis signal-regulating kinases (ASK1-3) are MAP3Ks that trigger cellular responses to redox stress and inflammatory cytokines (6, 7) and play vital roles in innate immunity and viral infection (8-11). When activated, ASK1-3 activate JNK and p38 via phosphorylation of MAP2Ks (MKK3/4/6/7) (12). The key initiator role of ASK1-3 in this pathway means that either too much or too little ASK activity can have pathological effects. For instance, inhibiting ASK1 is beneficial against gastric cancer (13,14), but inactivating mutations in ASK1...

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Assembly Dynamics and Stoichiometry of the Apoptosis Signal-regulating Kinase (ASK) Signalosome in Response to Electrophile Stress

Cited by 33 publications

References 69 publications

Mechanism of preferential complex formation by Apoptosis Signal-regulating Kinases

Mechanism of preferential complex formation by Apoptosis Signal-regulating Kinases

Characterization of the CLASP2 Protein Interaction Network Identifies SOGA1 as a Microtubule-Associated Protein

Structural basis of autoregulatory scaffolding by apoptosis signal-regulating kinase 1

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