PUBLICATION

Dynamics of in vivo ASC speck formation

Authors
Kuri, P., Schieber, N.L., Thumberger, T., Wittbrodt, J., Schwab, Y., Leptin, M.
ID
ZDB-PUB-170714-13
Date
2017
Source
The Journal of cell biology   216(9): 2891-2909 (Journal)
Registered Authors
Kuri, Paola, Leptin, Maria, Wittbrodt, Jochen
Keywords
none
MeSH Terms
  • Animals, Genetically Modified
  • Inflammasomes/drug effects
  • Inflammasomes/genetics
  • Inflammasomes/metabolism*
  • Inflammasomes/ultrastructure
  • NLR Proteins/genetics
  • NLR Proteins/metabolism
  • Mutation
  • Protein Interaction Domains and Motifs
  • Signal Transduction
  • Zebrafish/embryology
  • Zebrafish/genetics
  • Zebrafish/metabolism*
  • Phosphorylation
  • Microscopy, Electron, Transmission
  • Keratinocytes/drug effects
  • Keratinocytes/metabolism*
  • Keratinocytes/pathology
  • Keratinocytes/ultrastructure
  • Genotype
  • Pyroptosis
  • Time Factors
  • Protein Binding
  • Animals
  • Phenotype
  • Microscopy, Fluorescence
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
  • Zebrafish Proteins/ultrastructure
  • Microscopy, Video
  • CRISPR-Cas Systems
(all 31)
PubMed
28701426 Full text @ J. Cell Biol.
Abstract
Activated danger or pathogen sensors trigger assembly of the inflammasome adaptor ASC into specks, large signaling platforms considered hallmarks of inflammasome activation. Because a lack of in vivo tools has prevented the study of endogenous ASC dynamics, we generated a live ASC reporter through CRISPR/Cas9 tagging of the endogenous gene in zebrafish. We see strong ASC expression in the skin and other epithelia that act as barriers to insult. A toxic stimulus triggered speck formation and rapid pyroptosis in keratinocytes in vivo. Macrophages engulfed and digested that speck-containing, pyroptotic debris. A three-dimensional, ultrastructural reconstruction, based on correlative light and electron microscopy of the in vivo assembled specks revealed a compact network of highly intercrossed filaments, whereas pyrin domain (PYD) or caspase activation and recruitment domain alone formed filamentous aggregates. The effector caspase is recruited through PYD, whose overexpression induced pyroptosis but only after substantial delay. Therefore, formation of a single, compact speck and rapid cell-death induction in vivo requires a full-length ASC.
Errata / Notes
This article is corrected by ZDB-PUB-220906-82 .
Genes / Markers
Figures
No images available
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
hdb9Tgheat shockProtruding-mouth
hdb9Tgheat shockProtruding-mouth
hdb9Tgheat shockProtruding-mouth
hdb9Tgheat shock, chemical treatment by environment: apoptosis inhibitorProtruding-mouth
pycardhdb10Tgchemical treatment by environment: copper(II) sulfateProtruding-mouth
pycardhdb10Tgchemical treatment by environment: copper(II) sulfateProtruding-mouth
pycardhdb10Tgchemical treatment by environment: copper(II) sulfateProtruding-mouth
1 - 7 of 7
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
fr34TgTransgenic Insertion
    gl22TgTransgenic Insertion
      gz7TgTransgenic Insertion
        hdb2TgTransgenic Insertion
          hdb9TgTransgenic Insertion
            hdb10TgTransgenic Insertion
            hdb11
              		Small Deletion
              hdb12
                		Insertion
                nz50TgTransgenic Insertion
                  xt6TgTransgenic Insertion
                    1 - 10 of 11
                    Show
                    Human Disease / Model
                    Sequence Targeting Reagents
                    Fish
                    Antibodies
                    Orthology
                    Engineered Foreign Genes
                    Marker Marker Type Name
                    DsRed2EFGDsRed2
                    EGFPEFGEGFP
                    GAL4EFGGAL4
                    GFPEFGGFP
                    mKate2EFGmKate2
                    TagBFPEFGTagBFP
                    TagRFPEFGTagRFP
                    TomatoEFGTomato
                    1 - 8 of 8
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                    Mapping
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                    Citation
                    Kuri, P., Schieber, N.L., Thumberger, T., Wittbrodt, J., Schwab, Y., Leptin, M. (2017) Dynamics of in vivo ASC speck formation. The Journal of cell biology. 216(9):2891-2909.