CoChat
Sign Up
Cited by 14Cellular Mechanics and InteractionsCell Adhesion Molecules ResearchNF-κB Signaling Pathways

The DOI matches its Crossref record; 4 of the 4 references it cites were found in a registry or an index.

The Sharpin interactome reveals a role for Sharpin in lamellipodium formation via the Arp2/3 complex

Open access
Khan, Meraj H., Salomaa, Siiri I., Jacquemet, Guillaume, Butt, Umar, Miihkinen, Mitro, Deguchi, Takahiro, Kremneva, Elena, Lappalainen, Pekka, Humphries, Martin J., Pouwels, Jeroen·2017·Journal of Cell Science
DOI 10.1242/jcs.200329PMC PMC5612173
Open this paper in CoChat — free accountOpen PDF

Abstract

Sharpin, a multifunctional adaptor protein, regulates several signalling pathways. For example, Sharpin enhances signal-induced NF-κB signalling as part of the linear ubiquitin assembly complex (LUBAC) and inhibits integrins, the T cell receptor, caspase 1 and PTEN. However, despite recent insights into Sharpin and LUBAC function, a systematic approach to identify the signalling pathways regulated by Sharpin has not been reported. Here, we present the first 'Sharpin interactome', which identifies a large number of novel potential Sharpin interactors in addition to several known ones. These data suggest that Sharpin and LUBAC might regulate a larger number of biological processes than previously identified, such as endosomal trafficking, RNA processing, metabolism and cytoskeleton regulation. Importantly, using the Sharpin interactome, we have identified a novel role for Sharpin in lamellipodium formation. We demonstrate that Sharpin interacts with Arp2/3, a protein complex that catalyses actin filament branching. We have identified the Arp2/3-binding site in Sharpin and demonstrate using a specific Arp2/3-binding deficient mutant that the Sharpin-Arp2/3 interaction promotes lamellipodium formation in a LUBAC-independent fashion.This article has an associated First Person interview with the first author of the paper.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1242/jcs.200329

Cites

4 of 4 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • SHARPIN regulates cell adhesion and migration either by inhibition of integrins or by promotion of lamellipodium formation through the ARP2/3 complex.

    “What is known is that SHARPIN regulates cell adhesion and migration either by inhibition of integrins”

    From SHARPIN S146 phosphorylation mediates ARP2/3 interaction, cancer cell invasion and metastasis (2022) · cites this paper as Khan et al., 2017

    In ‘The Sharpin-Arp2/3 interaction promotes cell migration’ (page unavailable), the paper says Sharpin might regulate migration through integrin inhibition and Arp2/3-dependent lamellipodium formation. It also reports migration promoted by the Sharpin–Arp2/3 interaction in a specific cell assay. The claim presents both routes as established, whereas the paper hedges that broader conclusion and notes that their effects appeared to cancel in its wound-healing assay. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

  • ARP2/3 interaction with SHARPIN promotes lamellipodia formation.

    “We have previously shown that ARP2/3 interaction with SHARPIN promotes lamellipodia formation (Khan et al., 2017).”

    From SHARPIN S146 phosphorylation mediates ARP2/3 interaction, cancer cell invasion and metastasis (2022) · cites this paper as Khan et al., 2017

  • Prior to this study, SHARPIN was known to promote lamellipodium formation through interaction with ARP2/3 complex.

    “Prior this study, SHARPIN was known to promote lamellipodium formation through interaction with the ARP2/3 complex”

    From SHARPIN S146 phosphorylation mediates ARP2/3 interaction, cancer cell invasion and metastasis (2022) · cites this paper as Khan et al., 2017

  • The assays used for integrin activity, LUBAC activity, and ARP2/3 interaction were previously established.

    “As functional read-outs, we used previously established assays for three SHARPIN-regulated functions: integrin activity, LUBAC activity, and ARP2/3 interaction (Bouaouina, Harburger et al., 2011, Harburger, Bouaouina et al., 2009, Khan et al., 2017).”

    From SHARPIN S146 phosphorylation mediates ARP2/3 interaction, cancer cell invasion and metastasis (2022) · cites this paper as Khan et al., 2017

  • This function was independent of its LUBAC- and integrin related roles.

    “it was further demonstrated that this function was independent of its LUBAC- and integrin related roles (Khan et al., 2017).”

    From SHARPIN S146 phosphorylation mediates ARP2/3 interaction, cancer cell invasion and metastasis (2022) · cites this paper as Khan et al., 2017

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Flagged first, then by citations

Research works printed but not linked: 62

  1. Azioune, 2009, Simple and rapid process for single cell micro-patterning, Lab. Chip, 9, 1640, 10.1039/b821581m · DOI
  2. Barsnes, 2009, PRIDE converter: making proteomics data-sharing easy, Nat. Biotechnol., 27, 598, 10.1038/nbt0709-598 · DOI
  3. Beli, 2008, WAVE and Arp2/3 jointly inhibit filopodium formation by entering into a complex with mDia2, Nat. Cell Biol., 10, 849, 10.1038/ncb1745 · DOI
  4. Bii, 2015, A novel gammaretroviral shuttle vector insertional mutagenesis screen identifies SHARPIN as a breast cancer metastasis gene and prognostic biomarker, Oncotarget, 6, 39507, 10.18632/oncotarget.6232 · DOI · Paper page
  5. Boisson, 2015, Human HOIP and LUBAC deficiency underlies autoinflammation, immunodeficiency, amylopectinosis, and lymphangiectasia, J. Exp. Med., 212, 939, 10.1084/jem.20141130 · DOI
  6. Brayford, 2016, Tropomyosin promotes lamellipodial persistence by collaborating with Arp2/3 at the leading edge, Curr. Biol., 26, 1312, 10.1016/j.cub.2016.03.028 · DOI
  7. Byron, 2015, A proteomic approach reveals integrin activation state-dependent control of microtubule cortical targeting, Nat. Commun., 6, 6135, 10.1038/ncomms7135 · DOI
  8. Carlier, 2000, GRB2 links signaling to actin assembly by enhancing interaction of neural wiskott-aldrich syndrome protein (N-WASp) with actin-related protein (ARP2/3) complex, J. Biol. Chem., 275, 21946, 10.1074/jbc.M000687200 · DOI
  9. Carpenter, 2006, CellProfiler: Image analysis software for identifying and quantifying cell phenotypes, Genome Biol., 7, R100, 10.1186/gb-2006-7-10-r100 · DOI
  10. Chattopadhyay, 2016, Ubiquitination of the transcription factor IRF-3 activates RIPA, the apoptotic pathway that protects mice from viral pathogenesis, Immunity, 44, 1151, 10.1016/j.immuni.2016.04.009 · DOI
  11. De Franceschi, 2015, Mutually exclusive roles of SHARPIN in integrin inactivation and NF-κB signaling, PLoS ONE, 10, e0143423, 10.1371/journal.pone.0143423 · DOI · Paper page
  12. de Hoon, 2004, Open source clustering software, Bioinformatics, 20, 1453, 10.1093/bioinformatics/bth078 · DOI
  13. Dubois, 2014, A catalytic-independent role for the LUBAC in NF-kappaB activation upon antigen receptor engagement and in lymphoma cells, Blood, 123, 2199, 10.1182/blood-2013-05-504019 · DOI
  14. Gerlach, 2011, Linear ubiquitination prevents inflammation and regulates immune signalling, Nature, 471, 591, 10.1038/nature09816 · DOI · Paper page
  15. Gustafsson, 2016, Fast live-cell conventional fluorophore nanoscopy with ImageJ through super-resolution radial fluctuations, Nat. Commun., 7, 12471, 10.1038/ncomms12471 · DOI
  16. Haynes, 2015, GMFbeta controls branched actin content and lamellipodial retraction in fibroblasts, J. Cell Biol., 209, 803, 10.1083/jcb.201501094 · DOI
  17. He, 2010, Shank-interacting protein-like 1 promotes tumorigenesis via PTEN inhibition in human tumor cells, J. Clin. Invest., 120, 2094, 10.1172/JCI40778 · DOI
  18. da Huang, 2009, Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources, Nat. Protoc., 4, 44, 10.1038/nprot.2008.211 · DOI · Paper page
  19. Ikeda, 2011, SHARPIN forms a linear ubiquitin ligase complex regulating NF-kappaB activity and apoptosis, Nature, 471, 637, 10.1038/nature09814 · DOI · Paper page
  20. Jung, 2010, Newly identified tumor-associated role of human sharpin, Mol. Cell. Biochem., 340, 161, 10.1007/s11010-010-0413-x · DOI · Paper page
  21. Krause, 2014, Steering cell migration: Lamellipodium dynamics and the regulation of directional persistence, Nat. Rev. Mol. Cell Biol., 15, 577, 10.1038/nrm3861 · DOI
  22. Kumari, 2014, Sharpin prevents skin inflammation by inhibiting TNFR1-induced keratinocyte apoptosis, Elife, 3, 10.7554/eLife.03422 · DOI
  23. Lai, 2008, Arp2/3 complex interactions and actin network turnover in lamellipodia, EMBO J., 27, 982, 10.1038/emboj.2008.34 · DOI
  24. Landgraf, 2010, Sipl1 and Rbck1 are novel Eya1-binding proteins with a role in craniofacial development, Mol. Cell. Biol., 30, 5764, 10.1128/MCB.01645-09 · DOI
  25. Lewis, 2015, UBE2L3 polymorphism amplifies NF-kappaB activation and promotes plasma cell development, linking linear ubiquitination to multiple autoimmune diseases, Am. J. Hum. Genet., 96, 221, 10.1016/j.ajhg.2014.12.024 · DOI
  26. Li, 2015, SHARPIN overexpression induces tumorigenesis in human prostate cancer LNCaP, DU145 and PC-3 cells via NF-kappaB/ERK/Akt signaling pathway, Med. Oncol., 32, 444, 10.1007/s12032-014-0444-3 · DOI · Paper page
  27. Liu, 2013, Expression of the Arp2/3 complex in human gliomas and its role in the migration and invasion of glioma cells, Oncol. Rep., 30, 2127, 10.3892/or.2013.2669 · DOI · Paper page
  28. Merico, 2010, Enrichment map: a network-based method for gene-set enrichment visualization and interpretation, PLoS ONE, 5, e13984, 10.1371/journal.pone.0013984 · DOI · Paper page
  29. Nastase, 2016, An essential role for SHARPIN in the regulation of caspase 1 activity in sepsis, Am. J. Pathol., 186, 1206, 10.1016/j.ajpath.2015.12.026 · DOI
  30. Nolen, 2009, Characterization of two classes of small molecule inhibitors of Arp2/3 complex, Nature, 460, 1031, 10.1038/nature08231 · DOI
  31. Park, 2016, SHARPIN controls regulatory T cells by negatively modulating the T cell antigen receptor complex, Nat. Immunol., 17, 286, 10.1038/ni.3352 · DOI
  32. Perez-Riverol, 2016, PRIDE inspector toolsuite: Moving toward a universal visualization tool for proteomics data standard formats and quality assessment of ProteomeXchange datasets, Mol. Cell. Proteomics, 15, 305, 10.1074/mcp.O115.050229 · DOI
  33. Pollard, 1984, Quantitative analysis of the effect of acanthamoeba profilin on actin filament nucleation and elongation, Biochemistry, 23, 6631, 10.1021/bi00321a054 · DOI
  34. Pouwels, 2013, SHARPIN regulates uropod detachment in migrating lymphocytes, Cell. Rep., 5, 619, 10.1016/j.celrep.2013.10.011 · DOI · Paper page
  35. Prehoda, 2000, Integration of multiple signals through cooperative regulation of the N-WASP-Arp2/3 complex, Science, 290, 801, 10.1126/science.290.5492.801 · DOI
  36. Ran, 2013, Genome engineering using the CRISPR-Cas9 system, Nat. Protoc., 8, 2281, 10.1038/nprot.2013.143 · DOI · Paper page
  37. Rantala, 2011, SHARPIN is an endogenous inhibitor of beta1-integrin activation, Nat. Cell Biol., 13, 1315, 10.1038/ncb2340 · DOI · Paper page
  38. Rickard, 2014, TNFR1-dependent cell death drives inflammation in sharpin-deficient mice, Elife, 3, 10.7554/eLife.03464 · DOI
  39. Riedl, 2008, Lifeact: a versatile marker to visualize F-actin, Nat. Methods, 5, 605, 10.1038/nmeth.1220 · DOI · Paper page
  40. Rogers, 2003, Molecular requirements for actin-based lamella formation in drosophila S2 cells, J. Cell Biol., 162, 1079, 10.1083/jcb.200303023 · DOI
  41. Rohatgi, 1999, The interaction between N-WASP and the Arp2/3 complex links Cdc42-dependent signals to actin assembly, Cell, 97, 221, 10.1016/S0092-8674(00)80732-1 · DOI
  42. Rotty, 2013, New insights into the regulation and cellular functions of the ARP2/3 complex, Nat. Rev. Mol. Cell Biol., 14, 7, 10.1038/nrm3492 · DOI · Paper page
  43. Rozelle, 2000, Phosphatidylinositol 4,5-bisphosphate induces actin-based movement of raft-enriched vesicles through WASP-Arp2/3, Curr. Biol., 10, 311, 10.1016/S0960-9822(00)00384-5 · DOI
  44. Saldanha, 2004, Java treeview–extensible visualization of microarray data, Bioinformatics, 20, 3246, 10.1093/bioinformatics/bth349 · DOI
  45. Schindelin, 2012, Fiji: An open-source platform for biological-image analysis, Nat. Methods, 9, 676, 10.1038/nmeth.2019 · DOI · Paper page
  46. Shevchenko, 1996, A strategy for identifying gel-separated proteins in sequence databases by MS alone, Biochem. Soc. Trans., 24, 893, 10.1042/bst0240893 · DOI
  47. Smoot, 2011, Cytoscape 2.8: new features for data integration and network visualization, Bioinformatics, 27, 431, 10.1093/bioinformatics/btq675 · DOI
  48. Söderberg, 2006, Direct observation of individual endogenous protein complexes in situ by proximity ligation, Nat. Methods, 3, 995, 10.1038/nmeth947 · DOI · Paper page
  49. Stieglitz, 2012, Structural analysis of SHARPIN, a subunit of a large multi-protein E3 ubiquitin ligase, reveals a novel dimerization function for the pleckstrin homology superfold, J. Biol. Chem., 287, 20823, 10.1074/jbc.M112.359547 · DOI
  50. Subauste, 2000, Rho family proteins modulate rapid apoptosis induced by cytotoxic T lymphocytes and fas, J. Biol. Chem., 275, 9725, 10.1074/jbc.275.13.9725 · DOI
  51. Suraneni, 2012, The Arp2/3 complex is required for lamellipodia extension and directional fibroblast cell migration, J. Cell Biol., 197, 239, 10.1083/jcb.201112113 · DOI · Paper page
  52. Tokunaga, 2009, Involvement of linear polyubiquitylation of NEMO in NF-kappaB activation, Nat. Cell Biol., 11, 123, 10.1038/ncb1821 · DOI
  53. Vizcaíno, 2010, The proteomics identifications database: 2010 update, Nucleic Acids Res., 38, D736, 10.1093/nar/gkp964 · DOI
  54. Vizcaíno, 2013, The PRoteomics IDEntifications (PRIDE) database and associated tools: Status in 2013, Nucleic Acids Res., 41, D1063, 10.1093/nar/gks1262 · DOI · Paper page
  55. Wagner, 2016, SPATA2 links CYLD to the TNF-alpha receptor signaling complex and modulates the receptor signaling outcomes, EMBO J., 35, 1868, 10.15252/embj.201694300 · DOI
  56. Welch, 1997, The human Arp2/3 complex is composed of evolutionarily conserved subunits and is localized to cellular regions of dynamic actin filament assembly, J. Cell Biol., 138, 375, 10.1083/jcb.138.2.375 · DOI
  57. Wu, 2009, Integrated network analysis platform for protein-protein interactions, Nat. Methods, 6, 75, 10.1038/nmeth.1282 · DOI
  58. Wu, 2012, Arp2/3 is critical for lamellipodia and response to extracellular matrix cues but is dispensable for chemotaxis, Cell, 148, 973, 10.1016/j.cell.2011.12.034 · DOI
  59. Yang, 2014, Essential role of the linear ubiquitin chain assembly complex in lymphoma revealed by rare germline polymorphisms, Cancer. Discov., 4, 480, 10.1158/2159-8290.CD-13-0915 · DOI
  60. Yang, 2016, Targeting non-proteolytic protein ubiquitination for the treatment of diffuse large B cell lymphoma, Cancer. Cell., 29, 494, 10.1016/j.ccell.2016.03.006 · DOI
  61. Zak, 2011, Systems analysis identifies an essential role for SHANK-associated RH domain-interacting protein (SHARPIN) in macrophage toll-like receptor 2 (TLR2) responses, Proc. Natl. Acad. Sci. USA, 108, 11536, 10.1073/pnas.1107577108 · DOI
  62. Zhang, 2014, Activation of nuclear factor kappaB pathway and downstream targets survivin and livin by SHARPIN contributes to the progression and metastasis of prostate cancer, Cancer, 120, 3208, 10.1002/cncr.28796 · DOI

Cited by

Sorted by citations

Metadata

Authors
Khan, M. H., Salomaa, S. I., Jacquemet, G., Butt, U., Miihkinen, M., Deguchi, T., Kremneva, E., Lappalainen, P., Humphries, M. J., & Pouwels, J.
Affiliations
  • Åbo Akademi University (FI)
  • University of Turku (FI)
  • University of Helsinki (FI)
  • Wellcome Centre for Cell-Matrix Research (GB)
Corresponding author
Pouwels, Jeroen
ORCID
Venue
Journal of Cell Science
Publisher
The Company of Biologists
Published
2017-09-15
Volume
130
Issue
18
Pages
3094-3107
ISSN
  • 0021-9533 (ISSN-L)
  • 1477-9137
DOI
10.1242/jcs.200329
PMC
PMC5612173
OpenAlex
W2744896017
Cited by
14
References
4
Citations by year
2022: 2 · 2021: 2 · 2020: 6 · 2019: 2 · 2017: 1
FWCI
1.31
Type
journal article
Language
en
Open access
Licence
www.biologists.com/user-licence-1-1/ (vor, 181 d)
Locations
  • Journal of Cell Science (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • UTUPub (University of Turku) (submittedVersion)
  • HighWire Press Open Archive (submittedVersion)
  • Research Explorer (The University of Manchester) (submittedVersion)
  • Åbo Akademi University Research Portal (submittedVersion)
Funders
  • Turun Yliopisto
  • Sigrid Juséliuksen Säätiö
  • Wellcome Trust (092015)
  • Cancer Research UK (C13329/A21671)
  • Academy of Finland
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
Keywords
SHARPIN, protein-protein interaction network, Arp2/3 complex, lamellipodium formation, actin cytoskeleton, cell motility, endosomal trafficking, RNA processing, metabolism
MeSH terms
Cell Movement, Cytoskeleton, HeLa Cells, Humans, Nerve Tissue Proteins, Protein Binding, Pseudopodia, Mass Spectrometry, Actin-Related Protein 2-3 Complex, Time-Lapse Imaging, Protein Interaction Maps, Gene Ontology

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review