Proteolytic processing of nuclear factor kappa B by calpain in vitro.
Nuclear factor kappaB (NF-kappaB) is a transcription factor that is critical for the inducible expression of multiple cellular and viral genes. Using the electrophoretic mobility shift assay, we demonstrated that DNA binding activity of NF-kappaB was abolished by proteolysis with mu- and m-calpains in vitro. The proteolysis of NF-kappaB by calpains and hence the abolition of its DNA binding was prevented by calpastatin, calpain inhibitor I and proteasome inhibitor. We also provided evidence that calpains degrade the C-terminal domain of NF-kappaB by Western blot using anti-NF-kappaB (p65) C-terminal antibody. These observations indicate that calpains regulate gene expression through processing of NF-kappaB.
Liu
Z Q
ZQ
Department of Molecular Genetics, Nagoya City University Medical School, Japan.
Kunimatsu
M
M
Yang
J P
JP
Ozaki
Y
Y
Sasaki
M
M
Okamoto
T
T
D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000494:pharmacology; D002154:Calpain / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004247:DNA / Q000378:metabolism; D006023:Glycoproteins / Q000494:pharmacology; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000302:isolation & purification / Q000378:metabolism*; D011499:Protein Processing, Post-Translational
D002135:Calcium-Binding Proteins; D015853:Cysteine Proteinase Inhibitors; D006023:Glycoproteins; D007976:Leupeptins; D016328:NF-kappa B; C039304:calpain inhibitors; C089486:carbobenzoxy-leucyl-leucyl-norvalinal; C033668:calpastatin; D004247:DNA; D002154:Calpain