Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Yin, Chao, Gu, Jiaojie, Gu, Dan, Wang, Zhenyu, Ji, Ruoyun, Jiao, Xinan, Li, Qiuchun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
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
_version_ 1784845362702319616
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
work_keys_str_mv AT yinchao thesalmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT gujiaojie thesalmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT gudan thesalmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT wangzhenyu thesalmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT jiruoyun thesalmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT jiaoxinan thesalmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT liqiuchun thesalmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT yinchao salmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT gujiaojie salmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT gudan salmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT wangzhenyu salmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT jiruoyun salmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT jiaoxinan salmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway
AT liqiuchun salmonellat3ss1effectoripajisregulatedbyitraandinhibitsthemapksignalingpathway