Intracellular trafficking of AIP56, an NF-κB-cleaving toxin from Photobacterium damselae subsp. Piscicida
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10216/114494 |
Resumo: | AIP56 (apoptosis-inducing protein of 56 kDa) is a metalloprotease AB toxin secreted by Photobacterium damselae subsp. piscicida that acts by cleaving NF-κB. During infection, AIP56 spreads systemically and depletes phagocytes by postapoptotic secondary necrosis, impairing the host phagocytic defense and contributing to the genesis of infection-associated necrotic lesions. Here we show that mouse bone marrow-derived macrophages (mBMDM) intoxicated by AIP56 undergo NF-κB p65 depletion and apoptosis. Similarly to what was reported for sea bass phagocytes, intoxication of mBMDM involves interaction of AIP56 C-terminal region with cell surface components, suggesting the existence of a conserved receptor. Biochemical approaches and confocal microscopy revealed that AIP56 undergoes clathrin-dependent endocytosis, reaches early endosomes, and follows the recycling pathway. Translocation of AIP56 into the cytosol requires endosome acidification, and an acidic pulse triggers translocation of cell surface-bound AIP56 into the cytosol. Accordingly, at acidic pH, AIP56 becomes more hydrophobic, interacting with artificial lipid bilayer membranes. Altogether, these data indicate that AIP56 is a short-trip toxin that reaches the cytosol using an acidic-pH-dependent mechanism, probably from early endosomes. Usually, for short-trip AB toxins, a minor pool reaches the cytosol by translocating from endosomes, whereas the rest is routed to lysosomes for degradation. Here we demonstrate that part of endocytosed AIP56 is recycled back and released extracellularly through a mechanism requiring phosphoinositide 3-kinase (PI3K) activity but independent of endosome acidification. So far, we have been unable to detect biological activity of recycled AIP56, thereby bringing into question its biological relevance as well as the importance of the recycling pathway. |
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Intracellular trafficking of AIP56, an NF-κB-cleaving toxin from Photobacterium damselae subsp. PiscicidaAnimalsApoptosisApoptosis Regulatory Proteins/metabolismBacterial Toxins/metabolismCell SurvivalCells, CulturedCytosol/chemistryEndocytosisEndosomes/chemistryHydrogen-Ion ConcentrationMacrophages/microbiologyMacrophages/physiologyMaleMice, Inbred C57BLMicroscopy, ConfocalNF-kappa B/metabolismPeptide Hydrolases/metabolismPhotobacterium/metabolismProtein TransportProteolysisAIP56 (apoptosis-inducing protein of 56 kDa) is a metalloprotease AB toxin secreted by Photobacterium damselae subsp. piscicida that acts by cleaving NF-κB. During infection, AIP56 spreads systemically and depletes phagocytes by postapoptotic secondary necrosis, impairing the host phagocytic defense and contributing to the genesis of infection-associated necrotic lesions. Here we show that mouse bone marrow-derived macrophages (mBMDM) intoxicated by AIP56 undergo NF-κB p65 depletion and apoptosis. Similarly to what was reported for sea bass phagocytes, intoxication of mBMDM involves interaction of AIP56 C-terminal region with cell surface components, suggesting the existence of a conserved receptor. Biochemical approaches and confocal microscopy revealed that AIP56 undergoes clathrin-dependent endocytosis, reaches early endosomes, and follows the recycling pathway. Translocation of AIP56 into the cytosol requires endosome acidification, and an acidic pulse triggers translocation of cell surface-bound AIP56 into the cytosol. Accordingly, at acidic pH, AIP56 becomes more hydrophobic, interacting with artificial lipid bilayer membranes. Altogether, these data indicate that AIP56 is a short-trip toxin that reaches the cytosol using an acidic-pH-dependent mechanism, probably from early endosomes. Usually, for short-trip AB toxins, a minor pool reaches the cytosol by translocating from endosomes, whereas the rest is routed to lysosomes for degradation. Here we demonstrate that part of endocytosed AIP56 is recycled back and released extracellularly through a mechanism requiring phosphoinositide 3-kinase (PI3K) activity but independent of endosome acidification. So far, we have been unable to detect biological activity of recycled AIP56, thereby bringing into question its biological relevance as well as the importance of the recycling pathway.American Society for Microbiology20142014-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10216/114494eng0019-956710.1128/IAI.02623-14Pereira, LMPinto, RDSilva, DSMoreira, ARBeitzinger, COliveira, PSampaio, PBenz, RAzevedo, JESantos, NMVale, Ainfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-29T13:41:58Zoai:repositorio-aberto.up.pt:10216/114494Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T23:45:58.218850Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Intracellular trafficking of AIP56, an NF-κB-cleaving toxin from Photobacterium damselae subsp. Piscicida |
title |
Intracellular trafficking of AIP56, an NF-κB-cleaving toxin from Photobacterium damselae subsp. Piscicida |
spellingShingle |
Intracellular trafficking of AIP56, an NF-κB-cleaving toxin from Photobacterium damselae subsp. Piscicida Pereira, LM Animals Apoptosis Apoptosis Regulatory Proteins/metabolism Bacterial Toxins/metabolism Cell Survival Cells, Cultured Cytosol/chemistry Endocytosis Endosomes/chemistry Hydrogen-Ion Concentration Macrophages/microbiology Macrophages/physiology Male Mice, Inbred C57BL Microscopy, Confocal NF-kappa B/metabolism Peptide Hydrolases/metabolism Photobacterium/metabolism Protein Transport Proteolysis |
title_short |
Intracellular trafficking of AIP56, an NF-κB-cleaving toxin from Photobacterium damselae subsp. Piscicida |
title_full |
Intracellular trafficking of AIP56, an NF-κB-cleaving toxin from Photobacterium damselae subsp. Piscicida |
title_fullStr |
Intracellular trafficking of AIP56, an NF-κB-cleaving toxin from Photobacterium damselae subsp. Piscicida |
title_full_unstemmed |
Intracellular trafficking of AIP56, an NF-κB-cleaving toxin from Photobacterium damselae subsp. Piscicida |
title_sort |
Intracellular trafficking of AIP56, an NF-κB-cleaving toxin from Photobacterium damselae subsp. Piscicida |
author |
Pereira, LM |
author_facet |
Pereira, LM Pinto, RD Silva, DS Moreira, AR Beitzinger, C Oliveira, P Sampaio, P Benz, R Azevedo, JE Santos, NM Vale, A |
author_role |
author |
author2 |
Pinto, RD Silva, DS Moreira, AR Beitzinger, C Oliveira, P Sampaio, P Benz, R Azevedo, JE Santos, NM Vale, A |
author2_role |
author author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Pereira, LM Pinto, RD Silva, DS Moreira, AR Beitzinger, C Oliveira, P Sampaio, P Benz, R Azevedo, JE Santos, NM Vale, A |
dc.subject.por.fl_str_mv |
Animals Apoptosis Apoptosis Regulatory Proteins/metabolism Bacterial Toxins/metabolism Cell Survival Cells, Cultured Cytosol/chemistry Endocytosis Endosomes/chemistry Hydrogen-Ion Concentration Macrophages/microbiology Macrophages/physiology Male Mice, Inbred C57BL Microscopy, Confocal NF-kappa B/metabolism Peptide Hydrolases/metabolism Photobacterium/metabolism Protein Transport Proteolysis |
topic |
Animals Apoptosis Apoptosis Regulatory Proteins/metabolism Bacterial Toxins/metabolism Cell Survival Cells, Cultured Cytosol/chemistry Endocytosis Endosomes/chemistry Hydrogen-Ion Concentration Macrophages/microbiology Macrophages/physiology Male Mice, Inbred C57BL Microscopy, Confocal NF-kappa B/metabolism Peptide Hydrolases/metabolism Photobacterium/metabolism Protein Transport Proteolysis |
description |
AIP56 (apoptosis-inducing protein of 56 kDa) is a metalloprotease AB toxin secreted by Photobacterium damselae subsp. piscicida that acts by cleaving NF-κB. During infection, AIP56 spreads systemically and depletes phagocytes by postapoptotic secondary necrosis, impairing the host phagocytic defense and contributing to the genesis of infection-associated necrotic lesions. Here we show that mouse bone marrow-derived macrophages (mBMDM) intoxicated by AIP56 undergo NF-κB p65 depletion and apoptosis. Similarly to what was reported for sea bass phagocytes, intoxication of mBMDM involves interaction of AIP56 C-terminal region with cell surface components, suggesting the existence of a conserved receptor. Biochemical approaches and confocal microscopy revealed that AIP56 undergoes clathrin-dependent endocytosis, reaches early endosomes, and follows the recycling pathway. Translocation of AIP56 into the cytosol requires endosome acidification, and an acidic pulse triggers translocation of cell surface-bound AIP56 into the cytosol. Accordingly, at acidic pH, AIP56 becomes more hydrophobic, interacting with artificial lipid bilayer membranes. Altogether, these data indicate that AIP56 is a short-trip toxin that reaches the cytosol using an acidic-pH-dependent mechanism, probably from early endosomes. Usually, for short-trip AB toxins, a minor pool reaches the cytosol by translocating from endosomes, whereas the rest is routed to lysosomes for degradation. Here we demonstrate that part of endocytosed AIP56 is recycled back and released extracellularly through a mechanism requiring phosphoinositide 3-kinase (PI3K) activity but independent of endosome acidification. So far, we have been unable to detect biological activity of recycled AIP56, thereby bringing into question its biological relevance as well as the importance of the recycling pathway. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014 2014-01-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10216/114494 |
url |
http://hdl.handle.net/10216/114494 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
0019-9567 10.1128/IAI.02623-14 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
American Society for Microbiology |
publisher.none.fl_str_mv |
American Society for Microbiology |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799135776799195136 |