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The Salmonella T3SS1 effector IpaJ is regulated by ItrA and inhibits the MAPK signaling pathway
Invasion plasmid antigen J (IpaJ) is a protein with cysteine protease activity that is present in Salmonella and Shigella species. Salmonella enterica serovar Pullorum uses IpaJ to inhibit the NF-κB pathway and the subsequent inflammatory response, resulting in bacterial survival in host macrophages...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Public Library of Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728880/ https://www.ncbi.nlm.nih.gov/pubmed/36477497 http://dx.doi.org/10.1371/journal.ppat.1011005 |
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author | Yin, Chao Gu, Jiaojie Gu, Dan Wang, Zhenyu Ji, Ruoyun Jiao, Xinan Li, Qiuchun |
author_facet | Yin, Chao Gu, Jiaojie Gu, Dan Wang, Zhenyu Ji, Ruoyun Jiao, Xinan Li, Qiuchun |
author_sort | Yin, Chao |
collection | PubMed |
description | Invasion plasmid antigen J (IpaJ) is a protein with cysteine protease activity that is present in Salmonella and Shigella species. Salmonella enterica serovar Pullorum uses IpaJ to inhibit the NF-κB pathway and the subsequent inflammatory response, resulting in bacterial survival in host macrophages. In the present study, we performed a DNA pull-down assay and EMSA and identified ItrA, a new DeoR family transcriptional regulator that could control the expression of IpaJ by directly binding to the promoter of ipaJ. The deletion of itrA inhibited the transcription of ipaJ in Salmonella. Tn-Seq revealed that two regulators of Salmonella pathogenicity island 1 (SPI-1), namely HilA and HilD, regulated the secretion of IpaJ. The deletion of hilA, hilD or SPI-1 inhibited the secretion of IpaJ in both cultured medium and Salmonella-infected cells. In contrast, the strain with the deletion of ssrB (an SPI-2 regulator-encoding gene) displayed normal IpaJ secretion, indicating that IpaJ is an effector of the SPI-1-encoded type III secretion system (T3SS1). To further demonstrate the role of IpaJ in host cells, we performed quantitative phosphoproteomics and compared the fold changes in signaling molecules in HeLa cells infected with wild-type S. Pullorum C79-13 with those in HeLa cells infected with the ipaJ-deleted strain C79-13ΔpSPI12. Both phosphoproteomics and Western blot analyses revealed that p-MEK and p-ERK molecules were increased in C79-13ΔpSPI12- and C79-13ΔpSPI12-pipaJ(C45A)-infected cells; and Co-IP assays demonstrated that IpaJ interacts with Ras to reduce its ubiquitination, indicating that IpaJ can inhibit the activation of the MAPK signaling pathway. |
format | Online Article Text |
id | pubmed-9728880 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-97288802022-12-08 The Salmonella T3SS1 effector IpaJ is regulated by ItrA and inhibits the MAPK signaling pathway Yin, Chao Gu, Jiaojie Gu, Dan Wang, Zhenyu Ji, Ruoyun Jiao, Xinan Li, Qiuchun PLoS Pathog Research Article Invasion plasmid antigen J (IpaJ) is a protein with cysteine protease activity that is present in Salmonella and Shigella species. Salmonella enterica serovar Pullorum uses IpaJ to inhibit the NF-κB pathway and the subsequent inflammatory response, resulting in bacterial survival in host macrophages. In the present study, we performed a DNA pull-down assay and EMSA and identified ItrA, a new DeoR family transcriptional regulator that could control the expression of IpaJ by directly binding to the promoter of ipaJ. The deletion of itrA inhibited the transcription of ipaJ in Salmonella. Tn-Seq revealed that two regulators of Salmonella pathogenicity island 1 (SPI-1), namely HilA and HilD, regulated the secretion of IpaJ. The deletion of hilA, hilD or SPI-1 inhibited the secretion of IpaJ in both cultured medium and Salmonella-infected cells. In contrast, the strain with the deletion of ssrB (an SPI-2 regulator-encoding gene) displayed normal IpaJ secretion, indicating that IpaJ is an effector of the SPI-1-encoded type III secretion system (T3SS1). To further demonstrate the role of IpaJ in host cells, we performed quantitative phosphoproteomics and compared the fold changes in signaling molecules in HeLa cells infected with wild-type S. Pullorum C79-13 with those in HeLa cells infected with the ipaJ-deleted strain C79-13ΔpSPI12. Both phosphoproteomics and Western blot analyses revealed that p-MEK and p-ERK molecules were increased in C79-13ΔpSPI12- and C79-13ΔpSPI12-pipaJ(C45A)-infected cells; and Co-IP assays demonstrated that IpaJ interacts with Ras to reduce its ubiquitination, indicating that IpaJ can inhibit the activation of the MAPK signaling pathway. Public Library of Science 2022-12-07 /pmc/articles/PMC9728880/ /pubmed/36477497 http://dx.doi.org/10.1371/journal.ppat.1011005 Text en © 2022 Yin et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Yin, Chao Gu, Jiaojie Gu, Dan Wang, Zhenyu Ji, Ruoyun Jiao, Xinan Li, Qiuchun The Salmonella T3SS1 effector IpaJ is regulated by ItrA and inhibits the MAPK signaling pathway |
title | The Salmonella T3SS1 effector IpaJ is regulated by ItrA and inhibits the MAPK signaling pathway |
title_full | The Salmonella T3SS1 effector IpaJ is regulated by ItrA and inhibits the MAPK signaling pathway |
title_fullStr | The Salmonella T3SS1 effector IpaJ is regulated by ItrA and inhibits the MAPK signaling pathway |
title_full_unstemmed | The Salmonella T3SS1 effector IpaJ is regulated by ItrA and inhibits the MAPK signaling pathway |
title_short | The Salmonella T3SS1 effector IpaJ is regulated by ItrA and inhibits the MAPK signaling pathway |
title_sort | salmonella t3ss1 effector ipaj is regulated by itra and inhibits the mapk signaling pathway |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728880/ https://www.ncbi.nlm.nih.gov/pubmed/36477497 http://dx.doi.org/10.1371/journal.ppat.1011005 |
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