title: Neuroepithelial transforming gene 1 (Net1) binds to caspase activation and recruitment domain (CARD)- and membrane-associated guanylate kinase-like domain-containing (CARMA) proteins and regulates nuclear factor κB activation.
abstract: The molecular complexes containing CARMA proteins have been recently identified as a key components in the signal transduction pathways that regulate activation of nuclear factor κB (NF-κB) transcription factor. Here, we used immunoprecipitation coupled with mass spectrometry to identify cellular binding partners of CARMA proteins. Our data indicate that the Rho guanine nucleotide exchange factor Net1 binds to CARMA1 and CARMA3 in resting and activated cells. Net1 expression induces NF-κB activation and cooperates with BCL10 and CARMA proteins in inducing NF-κB activity. Conversely, shRNA-mediated abrogation of Net1 results in impaired NF-κB activation following stimuli that require correct CARMA-BCL10-MALT1 complex formation and functioning. Microarray expression data are consistent with a positive role for Net1 on NF-κB activation. Thus, this study identifies Net1 as a CARMA-interacting molecule and brings important information on the molecular mechanisms that control NF-κB transcriptional activity.
authors.affiliation: Dipartimento di Scienze Biologiche ed Ambientali, Università degli Studi del Sannio, Via Port'Arsa 11, 82100 Benevento, Italy.
authors.forename: Mariangela
authors.initials: M
authors.lastname: Vessichelli
authors.forename: Angela
authors.initials: A
authors.lastname: Ferravante
authors.forename: Tiziana
authors.initials: T
authors.lastname: Zotti
authors.forename: Carla
authors.initials: C
authors.lastname: Reale
authors.forename: Ivan
authors.initials: I
authors.lastname: Scudiero
authors.forename: Gianluca
authors.initials: G
authors.lastname: Picariello
authors.forename: Pasquale
authors.initials: P
authors.lastname: Vito
authors.forename: Romania
authors.initials: R
authors.lastname: Stilo
mesh_terms: D053468:CARD Signaling Adaptor Proteins / Q000737:chemistry; D020169:Caspases / Q000378:metabolism*; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D019169:Jurkat Cells; D007963:Leukocytes, Mononuclear / Q000166:cytology; D000072000:NF-KappaB Inhibitor alpha; D020411:Oligonucleotide Array Sequence Analysis; D015513:Oncogene Proteins / Q000378:metabolism*; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry; D015398:Signal Transduction; D014162:Transfection
chemical_list: D053468:CARD Signaling Adaptor Proteins; D020672:I-kappa B Proteins; C099763:NET1 protein, human; C000606204:NFKBIA protein, human; D015513:Oncogene Proteins; D000072000:NF-KappaB Inhibitor alpha; D012313:RNA; D020169:Caspases