title: Specific effects of BCL10 Serine mutations on phosphorylations in canonical and noncanonical pathways of NF-κB activation following carrageenan.
abstract: To determine the impact of B cell leukemia/lymphoma (BCL) 10 on the phosphorylation of crucial mediators in NF-κB-mediated inflammatory pathways, human colonic epithelial cells were exposed to carrageenan (CGN), a sulfated polysaccharide commonly used as a food additive and known to induce NF-κB nuclear translocation by both canonical and noncanonical pathways. Phosphorylations of intermediates in inflammatory cascades, including NF-κB-inducing kinase (NIK) at Thr(559), transforming growth factor-β-activating kinase (TAK) 1 at Thr(184), Thr(187), and Ser(192), and inhibitory factor κBα (IκBα) at Ser(32), were examined following mutation of BCL10 at Ser(138) and at Ser(218). Specific phosphoantibodies were used for detection by enzyme-linked immunosorbent assay, immunoblot, and confocal microscopy of differences in phosphorylation following transfection by mutated BCL10. Both mutations demonstrated dominant-negative effects, with inhibition of phospho(Ser(32))-IκBα to less than control levels. Both of the BCL10 mutations reduced the CGN-induced increases in nuclear RelA and p50, but only the Ser(138) mutation inhibited the CGN-induced increases in nuclear RelB and p52 and in NIK Thr(559) phosphorylation. Hence, the phosphorylation of BCL10 Ser(138), but not Ser(218), emerged as a critical event in activation of the noncanonical pathway of NF-κB activation. Either BCL10 Ser(138) or Ser(218) mutation inhibited the phosphorylation of TAK1 at Thr(184) and at Thr(187), but not at Ser(192). These findings indicate that BCL10 phosphorylations act upstream of phosphorylations of NIK, TAK1, and IκBα and differentially affect the canonical and noncanonical pathways of NF-κB activation.
authors.affiliation: Department of Medicine, University of Illinois at Chicago, Chicago, Illinois 60612, USA.
authors.forename: Sumit
authors.initials: S
authors.lastname: Bhattacharyya
authors.forename: Alip
authors.initials: A
authors.lastname: Borthakur
authors.forename: Arivarasu N
authors.initials: AN
authors.lastname: Anbazhagan
authors.forename: Shivani
authors.initials: S
authors.lastname: Katyal
authors.forename: Pradeep K
authors.initials: PK
authors.lastname: Dudeja
authors.forename: Joanne K
authors.initials: JK
authors.lastname: Tobacman
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics*; D000074018:B-Cell CLL-Lymphoma 10 Protein; D001596:Benzylamines; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D002351:Carrageenan / Q000494:pharmacology; D002460:Cell Line; D003106:Colon / Q000166:cytology; D004847:Epithelial Cells / Q000187:drug effects; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism; D020930:MAP Kinase Kinase Kinases / Q000378:metabolism; D008297:Male; D018613:Microscopy, Confocal; D008875:Middle Aged; D016328:NF-kappa B / Q000187:drug effects / Q000378:metabolism*; D010766:Phosphorylation; D012694:Serine / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013449:Sulfonamides; D000099151:MAP Kinase Kinase Kinase 7; D000096926:Benzenesulfonamides
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000074018:B-Cell CLL-Lymphoma 10 Protein; D001596:Benzylamines; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002351:Carrageenan; D051550:I-kappa B Kinase; D020930:MAP Kinase Kinase Kinases; D016328:NF-kappa B; D012694:Serine; D013449:Sulfonamides; D000099151:MAP Kinase Kinase Kinase 7; C118591:BCL10 protein, human; C072105:KN 93; D000096926:Benzenesulfonamides