The serine/threonine phosphatase inhibitor calyculin A induces rapid degradation of IkappaBbeta. Requirement of both the N- and C-terminal sequences.
Signal-initiated activation of the transcription factor NF-kappaB is mediated through proteolysis of its cytoplasmic inhibitory proteins IkappaBalpha and IkappaBbeta. While most NF-kappaB inducers trigger the degradation of IkappaBalpha, only certain stimuli are able to induce the degradation of IkappaBbeta. The degradation of IkappaBalpha is targeted by its site-specific phosphorylations, although the mechanism underlying the degradation of IkappaBbeta remains elusive. In the present study, we have analyzed the effect of phosphatase inhibitors on the proteolysis of IkappaBbeta. We show that the serine/threonine phosphatase inhibitor calyculin A induces the hyperphosphorylation and subsequent degradation of IkappaBbeta in both human Jurkat T cells and the murine 70Z-3 preB cells, which is associated with the nuclear expression of active NF-kappaB. The calyculin A-mediated degradation of IkappaBbeta is further enhanced by the cytokine tumor necrosis factor-alpha (TNF-alpha), although TNF-alpha alone is unable to induce the degradation of IkappaBbeta. Mutational analyses have revealed that the inducible degradation of IkappaBbeta induced by calyculin A, and TNF-alpha requires two N-terminal serines (serines 19 and 23) that are homologous to the inducible phosphorylation sites present in IkappaBalpha. Furthermore, the C-terminal 51 amino acid residues, which are rich in serines and aspartic acids, are also required for the inducible degradation of IkappaBbeta. These results suggest that the degradation signal of IkappaBbeta may be controlled by the opposing actions of protein kinases and phosphatases and that both the N- and C-terminal sequences of IkappaBbeta are required for the inducible degradation of this NF-kappaB inhibitor.
Harhaj
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EW
Department of Microbiology and Immunology, Pennsylvania State University College of Medicine, Hershey Medical Center, Hershey, Pennsylvania 17033, USA.
Sun
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SC
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D020672:I-kappa B Proteins; D019169:Jurkat Cells; D008387:Marine Toxins; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D010080:Oxazoles / Q000494:pharmacology*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*; D010766:Phosphorylation; D013329:Structure-Activity Relationship; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; C083487:I kappa B beta protein; D020672:I-kappa B Proteins; D008387:Marine Toxins; D016328:NF-kappa B; D010080:Oxazoles; D014409:Tumor Necrosis Factor-alpha; C059041:calyculin A; D004247:DNA; D010749:Phosphoprotein Phosphatases