Porins from Salmonella enterica Serovar Typhimurium Activate the Transcription Factors Activating Protein 1 and NF-Œ∫B through the Raf-1-Mitogen-Activated Protein Kinase Cascade
Authors: Annalisa Tortora
Marina D&#x27;Isanto
Anna Longanella
Stefania Galdiero
Massimiliano Galdiero
Emma Sanzari
Mariateresa Vitiello
Journal: Infection and Immunity (2002)
Discipline: IMMUNOLOGY
Keywords: Salmonella enterica serovar Typhimurium; porins; AP-1; NF-kappaB; MAPK cascade
Affiliations: Istituto di Microbiologia, Facolt√† di Medicina e Chirurgia, Seconda Universit√† degli Studi di Napoli, 80138 Naples, Italy. , Istituto di Microbiologia, FacoltaÃÄ di Medicina e Chirurgia, Seconda UniversitaÃÄ degli Studi di Napoli, 80138 Naples, Dipartimento di Chimica Biologica, Centro di Studio di Biocristallografia, UniversitaÃÄ degli Studi di Napoli Federico II, 80134 Naples, Italy

Abstract:  In this study we examined the ability of Salmonella enterica serovar Typhimurium porins to activate activating protein 1 (AP-1) and nuclear factor kappaB (NF-kappaB) through the mitogen-activated protein kinase (MAPK) cascade, and we identified the AP-1-induced protein subunits. Our results demonstrate that these enzymes may participate in cell signaling pathways leading to AP-1 and NF-kappaB activation following porin stimulation of cells. Raf-1 was phosphorylated in response to the treatment of U937 cells with porins; moreover, the porin-mediated increase in Raf-1 phosphorylation is accompanied by the phosphorylation of MAPK kinase 1/2 (MEK1/2), p38, extracellular-signal-regulated kinase 1/2, and c-Jun N-terminal kinase. We used three different inhibitors of phosphorylation pathways: 2'-amino-3'-methoxyflavone (PD-098059), a selective inhibitor of MEK1 activator and the MAPK cascade; 4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole (SB203580), a specific inhibitor of the p38 pathway; and 7beta-acetoxy-1alpha,6beta,9alpha-trihydroxy-8,13-epoxy-labd-14-en-11-one (forskolin), an inhibitor at the level of Raf-1 kinase. PD-098059 pretreatment of cells decreases AP-1 and NF-kappaB activation by lipopolysaccharide (LPS) but not by porins, and SB203580 pretreatment of cells decreases mainly AP-1 and NF-kappaB activation by porins; in contrast, forskolin pretreatment of cells does not affect AP-1 and NF-kappaB activation following either porin or LPS stimulation. Our data suggest that the p38 signaling pathway mainly regulates AP-1 and NF-kappaB activation in cells treated with S. enterica serovar Typhimurium porins. Antibody electrophoretic mobility shift assays showed that JunD and c-Fos binding is found in cells treated with porins, in cells treated with LPS, and in unstimulated cells. However, by 30 to 60 min of stimulation, a different complex including c-Jun appears in cells treated with porins or LPS, while the Fra-2 subunit is present only after porin stimulation. These data suggest different molecular mechanisms of activation induced by porins or by LPS. 

Retracted: No

PubPeer comments:
 This paper is part of a set of at least 14 papers with shared authors (always including one with the last name Galdiero), spanning 1995-2008, that all appear to show the same gels. The porin gels appear to all represent the same sample (Salmonella typhimurium porins), but the LPS gels represent Salmonella, Pasteurella multocida, or Pasteurella haemolytica). Sometimes fewer or more lanes are visible, and in each paper the data are represented as novel. Here are the papers: * Mariateresa Vitiello et al., Immunology and Cell Biology (1995), DOI: 10.1038 icb.1995.70 - https: pubpeer.com publications 17EC2E40E190E2FCC6F181A2293361 * M. Galdiero et al., Immunology (1995), PMID: 8567029 - https: pubpeer.com publications DECE082D02578F84676293D3A3D307 * L. Sommese et al., J Med Microbiology (1996), DOI: 10.1099 00222615-45-1-40 - https: pubpeer.com publications 7DF0A919C0BB60C8DD14EA1AA8DF34 * M Galdiero et al., Immunology (1998), DOI: 10.1046 j.1365-2567.1998.00492.x - https: pubpeer.com publications BE8182500B61343C93C4D083288F5A * Giuseppe Iovane et al., Veterinary Immunology and Immunopathology (1998), DOI: 10.1016 S0165-2427(98)00183-4 - https: pubpeer.com publications 6B20332BB1333F6A9128F81ADE8047 * M. Galdiero et al., Clin Exp Immunol (1999), DOI: 10.1046 j.1365-2249.1999.00904.x - https: pubpeer.com publications 7489649618C468805E4E8A2DA683BB * Antonella Marcatili et al., Research in Microbiology (2000), DOI: 10.1016 S0923-2508(00)00142-X - https: pubpeer.com publications AE562BC01FEC2640B955C330A0B013 * M. Galdiero et al., Microbiology (2001), DOI: 10.1099 00221287-147-10-2697 - https: pubpeer.com publications 511A96912036B49FECC6CEB6F1CBF2 * Massimiliano Galdiero et al., Infection and Immunity (2002), DOI: 10.1128 iai.70.2.558-568.2002 - https: pubpeer.com publications A8765584CC85E943BCA5CB79901DAC * M. Galdiero et al., Journal of Infection (2003), DOI: 10.1053 jinf.2002.1068 - https: pubpeer.com publications FED37BABEA925C28B2CF7BF86CE5B8 * M. Galdiero et al., Clin Microbiol Infect (2003), DOI: 10.1046 j.1469-0691.2003.00728.x - https: pubpeer.com publications FC98EF7F5F87C7AE81F88F12D9C8F4 * M. Vitiello et al., Cytokine (2004), DOI: 10.1016 j.cyto.2004.03.010 - https: pubpeer.com publications 47A3D51AB3A1C226D75197C36D5753 * M. Galdiero et al., Med Microbiol Immunol (2005), DOI: 10.1007 s00430-003-0209-7 - https: pubpeer.com publications 850E4A39A93C3F04AC922264AFC923 * M. Vitiello et al., Cytokine (2008), DOI: 10.1016 j.cyto.2007.11.021 - https: pubpeer.com publications 1EB6BE6DC91E81C414E128642A2FA9 Here is a compilation of most of these gels, as made by Edentulina Martensi, for which I thank this PubPeer user.    , 
 Figures 2 and 3. * Boxes of the same color focus attention on lanes that appear to look similar across different panels and across the two figures.  , 
 Adding some more papers to the ones found by Dr. Bik and me. Dr. Bik:  This paper is part of a set of at least 14 papers with shared authors (always including one with the last name Galdiero), spanning 1995-2008, that all appear to show the same gels. The porin gels appear to all represent the same sample (Salmonella typhimurium porins), but the LPS gels represent Salmonella, Pasteurella multocida, or Pasteurella haemolytica). Sometimes fewer or more lanes are visible, and in each paper the data are represented as novel. Here is a compilation of most of these gels, as made by Edentulina Martensi, for which I thank this PubPeer user. Here are the papers: - Mariateresa Vitiello et al., Immunology and Cell Biology (1995), DOI: 10.1038 icb.1995.70 - https: pubpeer.com publications 17EC2E40E190E2FCC6F181A2293361 - M. Galdiero et al., Immunology (1995), PMID: 8567029 - https: pubpeer.com publications DECE082D02578F84676293D3A3D307 - L. Sommese et al., J Med Microbiology (1996), DOI: 10.1099 00222615-45-1-40 - https: pubpeer.com publications 7DF0A919C0BB60C8DD14EA1AA8DF34 - M Galdiero et al., Immunology (1998), DOI: 10.1046 j.1365-2567.1998.00492.x - https: pubpeer.com publications BE8182500B61343C93C4D083288F5A - Giuseppe Iovane et al., Veterinary Immunology and Immunopathology (1998), DOI: 10.1016 S0165-2427(98)00183-4 https: pubpeer.com publications 6B20332BB1333F6A9128F81ADE8047 - M. Galdiero et al., Clin Exp Immunol (1999), DOI: 10.1046 j.1365-2249.1999.00904.x - https: pubpeer.com publications 7489649618C468805E4E8A2DA683BB - Antonella Marcatili et al., Research in Microbiology (2000), DOI: 10.1016 S0923-2508(00)00142-X - https: pubpeer.com publications AE562BC01FEC2640B955C330A0B013 - M. Galdiero et al., Microbiology (2001), DOI: 10.1099 00221287-147-10-2697 - https: pubpeer.com publications 511A96912036B49FECC6CEB6F1CBF2 - Massimiliano Galdiero et al., Infection and Immunity (2002), DOI: 10.1128 iai.70.2.558-568.2002 - https: pubpeer.com publications A8765584CC85E943BCA5CB79901DAC - M. Galdiero et al., Journal of Infection (2003), DOI: 10.1053 jinf.2002.1068 - https: pubpeer.com publications FED37BABEA925C28B2CF7BF86CE5B8 - M. Galdiero et al., Clin Microbiol Infect (2003), DOI: 10.1046 j.1469-0691.2003.00728.x - https: pubpeer.com publications FC98EF7F5F87C7AE81F88F12D9C8F4 - M. Vitiello et al., Cytokine (2004), DOI: 10.1016 j.cyto.2004.03.010 - https: pubpeer.com publications 47A3D51AB3A1C226D75197C36D5753 - M. Galdiero et al., Med Microbiol Immunol (2005), DOI: 10.1007 s00430-003-0209-7 - https: pubpeer.com publications 850E4A39A93C3F04AC922264AFC923 - M. Vitiello et al., Cytokine (2008), DOI: 10.1016 j.cyto.2007.11.021 - https: pubpeer.com publications 1EB6BE6DC91E81C414E128642A2FA9     Here my edit: - M. Galdiero et al., Immunobiology (2000) DOI: 10.1016 S0171-2985(00)80030-3 https: pubpeer.com publications DBF917296422998A5EC5E6A50170A9 - G. Iovane et al., FEMS Immunology and Medical Microbiology (1998) DOI: 10.1111 j.1574-695x.1998.tb01108.x https: pubpeer.com publications 6A9702D24AB5F8283FCB210414880C - Marilena Galdiero et al., INFECTION AND IMMUNITY (2004), DOI: 10.1128 IAI.72.2.1204u20131209.2004 https: pubpeer.com publications 81F27E6CD12B2FCE200A53F1EDE8A9 In particular, in the lattest, of the rows in Figure 1 has a lot of similarities in common with another one in another paper , but there is one big difference (green arrow). Here compared:  ,