Proximity labeling of the Listeria monocytogenes surface reveals pathogen control of a host deubiquitinase

Abstract

ABSTRACT Intracellular pathogens must navigate the crowded cellular environment to establish infection. Listeria monocytogenes achieves this by recruiting host factors to its surface to hijack the host actin cytoskeleton for motility, form membrane protrusions, and spread from cell to cell. Although these types of Listeria -host interactions are critical for infection, systematic characterization of this interface has been limited. Here, we implement surface display of the promiscuous biotin ligase split-TurboID to profile host proteins recruited to the surface of L. monocytogenes during intracellular infection. This approach identified the host deubiquitinase CYLD as a protein selectively enriched at the pathogen surface. While CYLD promotes infection by suppressing autophagy and innate immunity in macrophages, how L. monocytogenes recruits and appropriates CYLD function in other cell types has remained unclear. We demonstrate that the E3 ligase RNF213 decorates bacterial poles with M1-linked linear ubiquitin, thereby redirecting CYLD to the bacterial surface. We further show ubiquitin is not sufficient to recruit CYLD but requires the L. monocytogenes secreted effector internalin C (InlC). Despite its presence at the bacterial surface, CYLD does not deubiquitinate bacteria or regulate autophagic bacterial clearance in infected epithelial cells. Instead, CYLD and InlC protect L. monocytogenes from IFN-γ- and RNF213-dependent restriction of cell-to-cell spread. Overall, our work profiling the bacterial surface-host interactome has identified a new mechanism by which InlC spatially reprograms CYLD activity, uncoupling its canonical immune functions to promote cell-to-cell spread in epithelial cells. These findings highlight how L. monocytogenes exploits, a host deubiquitinase, to perform cell-type-specific functions during infection.

The authors' abstract, as published at the source. Nature Communications, 2026 · DOI ↗