ACTIVATION INVITRO OF NF-KAPPA-B BY PHOSPHORYLATION OF ITS INHIBITOR I-KAPPA-B

被引:1176
作者
GHOSH, S [1 ]
BALTIMORE, D [1 ]
机构
[1] MIT,DEPT BIOL,CAMBRIDGE,MA 02139
关键词
D O I
10.1038/344678a0
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
NUCLEAR factor κB (NF-κB), which was first detected by its binding to the κB site in the immunoglobulin κ-gene enhancer 1, is important for the regulated expression of the κ-gene 2,3 and is partly responsible for the induction in appropriate cells of inter-leukin-2 (IL-2)4, IL-2α receptor5, β-interferon6,7 and serum amyloid A protein8. NF-κ is present as a nuclear DNA-binding protein in B lymphocytes and mature macrophages, but is found in the cytoplasm of many cells in a form unable to bind to DNA9. The cytoplasmic form is bound to an inhibitor protein, IκB, from which it can be released in vitro by deoxycholate and other agents10. Activation of cells by various agents, notably the phorbol esters that stimulate protein kinase C (PKC), leads to dissociation in vivo of the NF-κB/IκB complex and migration of NF-κB to the nucleus9. Therefore, it acts as a second messenger system, transducing activation signals from the cytoplasm to the nucleus. To elucidate the mechanism of signal transfer, we have used an in vitro system in which addition of purified protein kinases to a partially purified NF-κB/IκB complex leads to the activation of the DNA-binding activity of NF-κB. Using gel retardation assays we found that PKC, cyclic AMP-dependent protein kinase (PKA) and a haem-regulated eIF-2 kinase (HRI) could activate NF-κB in vitro, whereas casein kinase II was ineffective. To determine the target for the protein kinases we purified and characterized both NF-κB and IκB and found that IκB is phosphorylated and inactivated in the presence of PKC and HRI but not PKA. © 1990 Nature Publishing Group.
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页码:678 / 682
页数:5
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