Dataset Information

A comprehensive set of ER PDI family members supports the biogenesis of pro-inflammatory interleukin 12 family cytokines

ABSTRACT: Cytokines of the interleukin 12 (IL-12) family are assembled combinatorially from shared alpha and beta subunits. A common theme is that human IL-12 family alpha subunits remain incompletely structured in isolation until they pair with a designate beta subunit. Accordingly, chaperones need to support and control specific assembly processes. Here, we site-specifically introduce photo-crosslinking amino acids into the IL-12 and IL-23 alpha subunits (IL-12alpha and IL-23alpha). This allows to stabilize transient chaperone:client complexes for mass spectrometry and reveals a large set of ER chaperones to interact with IL-12alpha and IL-23alpha. Among those, we focus on protein disulfide isomerase (PDI) family members and reveal IL-12 family subunits to be clients of several ill-characterized PDIs. We find that different PDIs show selectivity for different cysteines in IL-12alpha and IL-23alpha. Despite this, PDI binding generally stabilizes unassembled IL-12alpha and IL-23alpha against degradation. In contrast, alpha:beta assembly appears robust and only multiple simultaneous PDI depletions reduce IL-12 secretion. Our comprehensive analysis of the IL-12/IL-23 chaperone machinery reveals a hitherto uncharacterized role for several PDIs in this process. This extends our understanding of how cells accomplish the task of specific protein assembly reactions for signaling processes. Furthermore, our findings show that cytokine secretion can be modulated by targeting specific ER chaperones.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Nina Bach

LAB HEAD: Matthias J. Feige

PROVIDER: PXD036463 | Pride | 2022-11-28

REPOSITORIES: Pride

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Publications

A comprehensive set of ER protein disulfide isomerase family members supports the biogenesis of proinflammatory interleukin 12 family cytokines.

Mideksa Yonatan G YG Aschenbrenner Isabel I Fux Anja A Kaylani Dinah D Weiß Caroline A M CAM Nguyen Tuan-Anh TA Bach Nina C NC Lang Kathrin K Sieber Stephan A SA Feige Matthias J MJ

The Journal of biological chemistry 20221104 12

Cytokines of the interleukin 12 (IL-12) family are assembled combinatorially from shared α and β subunits. A common theme is that human IL-12 family α subunits remain incompletely structured in isolation until they pair with a designate β subunit. Accordingly, chaperones need to support and control specific assembly processes. It remains incompletely understood, which chaperones are involved in IL-12 family biogenesis. Here, we site-specifically introduce photocrosslinking amino acids into the I ...[more]

PMID: 36336075

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Project description:A promising alternative to antibiotics for treatment of Staphylococcus aureus infections is photodynamic inactivation (PDI), which employs a photosensitizer (PS) that produces cytotoxic reactive oxygen species (ROS) when exposed to molecular oxygen and antimicrobial blue light in the spectrum of 400-470 nm. Although the precise mechanistic basis of PDI has not been defined, the formation of ROS and free radicals that oxidize a number of cellular targets, including membrane lipids, damage to proteins and nucleic acids result in inactivation of essential cellular functions and subsequent cell death. Because PDI is non-selective and affects multiple cellular targets, development of resistance or tolerance to PDI has been considered to be unlikely and attempts to induce S. aureus resistance or tolerance upon repeated sub-lethal doses of PDI have not succeeded. However, multiple aspects of PDI suggest that development of tolerance is highly probable, in particular when PDI is used for treatment of infections where the environment at the infection site prevents penetration of PDI at a level sufficient to cause death of all bacteria and with tolerant phenotypes emerging from the surviving bacteria. In this study, we sought to identify the mechanisms that contribute to PDI tolerance in S. aureus. S. aureus HG003 and the isogenic HG003DmutSL strain with defects in DNA mismatch repair were used to evaluate the response of S. aureus and the roles of DNA mismatch repair and gene regulatory networks to repeated sublethal doses of PDI. Global transcriptome and genome analyses were used in an agnostic approach to identify the underlying transcriptional responses and genetic adaptations that occur as a result of repeated PDI and contribute to PDI tolerance. Our results reveal multiple metabolic, transport and cell wall biogenesis pathways that contribute to PDI tolerance, and a S. aureus regulatory gene likely responsible for the adaptive transcriptional response to PDI.

Dataset Information

A comprehensive set of ER PDI family members supports the biogenesis of pro-inflammatory interleukin 12 family cytokines

Publications

A comprehensive set of ER protein disulfide isomerase family members supports the biogenesis of proinflammatory interleukin 12 family cytokines.

Similar Datasets

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