Heterologous Expression and Assembly of Human TLR Signaling Components in <i>Saccharomyces cerevisiae</i>

Toll-like receptor (TLR) signaling is key to detect pathogens and initiating inflammation. Ligand recognition triggers the assembly of supramolecular organizing centers (SMOCs) consisting of large complexes composed of multiple subunits. Building such signaling hubs relies on Toll Interleukin-1 Rece...

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Autores principales:	Julia María Coronas-Serna, Elba del Val, Jonathan C. Kagan, María Molina, Víctor J. Cid
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Saccharomyces cerevisiae</i> humanized yeast innate immunity ERMES MyD88 TIRAP Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/503beb84fd9c427ea3f553bde88ef07f
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spelling	oai:doaj.org-article:503beb84fd9c427ea3f553bde88ef07f2021-11-25T16:54:42ZHeterologous Expression and Assembly of Human TLR Signaling Components in <i>Saccharomyces cerevisiae</i>10.3390/biom111117372218-273Xhttps://doaj.org/article/503beb84fd9c427ea3f553bde88ef07f2021-11-01T00:00:00Zhttps://www.mdpi.com/2218-273X/11/11/1737https://doaj.org/toc/2218-273XToll-like receptor (TLR) signaling is key to detect pathogens and initiating inflammation. Ligand recognition triggers the assembly of supramolecular organizing centers (SMOCs) consisting of large complexes composed of multiple subunits. Building such signaling hubs relies on Toll Interleukin-1 Receptor (TIR) and Death Domain (DD) protein-protein interaction domains. We have expressed TIR domain-containing components of the human myddosome (TIRAP and MyD88) and triffosome (TRAM and TRIF) SMOCs in <i>Saccharomyces cerevisiae,</i> as a platform for their study. Interactions between the TLR4 TIR domain, TIRAP, and MyD88 were recapitulated in yeast. Human TIRAP decorated the yeast plasma membrane (PM), except for the bud neck, whereas MyD88 was found at cytoplasmic spots, which were consistent with endoplasmic reticulum (ER)-mitochondria junctions, as evidenced by co-localization with Mmm1 and Mdm34, components of the ER and Mitochondria Encounter Structures (ERMES). The formation of MyD88-TIRAP foci at the yeast PM was reinforced by co-expression of a membrane-bound TLR4 TIR domain. Mutations in essential residues of their TIR domains aborted MyD88 recruitment by TIRAP, but their respective subcellular localizations were unaltered. TRAM and TRIF, however, did not co-localize in yeast. TRAM assembled long PM-bound filaments that were disrupted by co-expression of the TLR4 TIR domain. Our results evidence that the yeast model can be exploited to study the interactions and subcellular localization of human SMOC components in vivo.Julia María Coronas-SernaElba del ValJonathan C. KaganMaría MolinaVíctor J. CidMDPI AGarticle<i>Saccharomyces cerevisiae</i>humanized yeastinnate immunityERMESMyD88TIRAPMicrobiologyQR1-502ENBiomolecules, Vol 11, Iss 1737, p 1737 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Saccharomyces cerevisiae</i> humanized yeast innate immunity ERMES MyD88 TIRAP Microbiology QR1-502
spellingShingle	<i>Saccharomyces cerevisiae</i> humanized yeast innate immunity ERMES MyD88 TIRAP Microbiology QR1-502 Julia María Coronas-Serna Elba del Val Jonathan C. Kagan María Molina Víctor J. Cid Heterologous Expression and Assembly of Human TLR Signaling Components in <i>Saccharomyces cerevisiae</i>
description	Toll-like receptor (TLR) signaling is key to detect pathogens and initiating inflammation. Ligand recognition triggers the assembly of supramolecular organizing centers (SMOCs) consisting of large complexes composed of multiple subunits. Building such signaling hubs relies on Toll Interleukin-1 Receptor (TIR) and Death Domain (DD) protein-protein interaction domains. We have expressed TIR domain-containing components of the human myddosome (TIRAP and MyD88) and triffosome (TRAM and TRIF) SMOCs in <i>Saccharomyces cerevisiae,</i> as a platform for their study. Interactions between the TLR4 TIR domain, TIRAP, and MyD88 were recapitulated in yeast. Human TIRAP decorated the yeast plasma membrane (PM), except for the bud neck, whereas MyD88 was found at cytoplasmic spots, which were consistent with endoplasmic reticulum (ER)-mitochondria junctions, as evidenced by co-localization with Mmm1 and Mdm34, components of the ER and Mitochondria Encounter Structures (ERMES). The formation of MyD88-TIRAP foci at the yeast PM was reinforced by co-expression of a membrane-bound TLR4 TIR domain. Mutations in essential residues of their TIR domains aborted MyD88 recruitment by TIRAP, but their respective subcellular localizations were unaltered. TRAM and TRIF, however, did not co-localize in yeast. TRAM assembled long PM-bound filaments that were disrupted by co-expression of the TLR4 TIR domain. Our results evidence that the yeast model can be exploited to study the interactions and subcellular localization of human SMOC components in vivo.
format	article
author	Julia María Coronas-Serna Elba del Val Jonathan C. Kagan María Molina Víctor J. Cid
author_facet	Julia María Coronas-Serna Elba del Val Jonathan C. Kagan María Molina Víctor J. Cid
author_sort	Julia María Coronas-Serna
title	Heterologous Expression and Assembly of Human TLR Signaling Components in <i>Saccharomyces cerevisiae</i>
title_short	Heterologous Expression and Assembly of Human TLR Signaling Components in <i>Saccharomyces cerevisiae</i>
title_full	Heterologous Expression and Assembly of Human TLR Signaling Components in <i>Saccharomyces cerevisiae</i>
title_fullStr	Heterologous Expression and Assembly of Human TLR Signaling Components in <i>Saccharomyces cerevisiae</i>
title_full_unstemmed	Heterologous Expression and Assembly of Human TLR Signaling Components in <i>Saccharomyces cerevisiae</i>
title_sort	heterologous expression and assembly of human tlr signaling components in <i>saccharomyces cerevisiae</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/503beb84fd9c427ea3f553bde88ef07f
work_keys_str_mv	AT juliamariacoronasserna heterologousexpressionandassemblyofhumantlrsignalingcomponentsinisaccharomycescerevisiaei AT elbadelval heterologousexpressionandassemblyofhumantlrsignalingcomponentsinisaccharomycescerevisiaei AT jonathanckagan heterologousexpressionandassemblyofhumantlrsignalingcomponentsinisaccharomycescerevisiaei AT mariamolina heterologousexpressionandassemblyofhumantlrsignalingcomponentsinisaccharomycescerevisiaei AT victorjcid heterologousexpressionandassemblyofhumantlrsignalingcomponentsinisaccharomycescerevisiaei
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Heterologous Expression and Assembly of Human TLR Signaling Components in <i>Saccharomyces cerevisiae</i>

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