Nanoscale Organization of the Endogenous ASC Speck

Overview

Journal iScience

Publisher Cell Press

Date 2023 Dec 4

PMID 38047065

Authors

Ivo M Gluck

Grusha Primal Mathias

Sebastian Strauss

Virgile Rat

Irene Gialdini

Thomas Sebastian Ebert

Che Stafford

Ganesh Agam

Suliana Manley

Veit Hornung

Ralf Jungmann

Christian Sieben

Don C Lamb

Affiliations

Soon will be listed here.

Abstract

The NLRP3 inflammasome is a central component of the innate immune system. Its activation leads to formation of the ASC speck, a supramolecular assembly of the inflammasome adaptor protein ASC. Different models, based on ASC overexpression, have been proposed for the structure of the ASC speck. Using dual-color 3D super-resolution imaging (dSTORM and DNA-PAINT), we visualized the ASC speck structure following NLRP3 inflammasome activation using endogenous ASC expression. A complete structure was only obtainable by labeling with both anti-ASC antibodies and nanobodies. The complex varies in diameter between ∼800 and 1000 nm, and is composed of a dense core with emerging filaments. Dual-color confocal fluorescence microscopy indicated that the ASC speck does not colocalize with the microtubule-organizing center at late time points after Nigericin stimulation. From super-resolution images of whole cells, the ASC specks were sorted into a pseudo-time sequence indicating that they become denser but not larger during formation.

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