Mutant human cells with constitutive activation of NF-κB
Mutant human cells with constitutive activation of NF-κB
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DOI：
10.1073/pnas.0306812101
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发表时间：
2004-01-06
影响因子：
11.1
通讯作者：
Stark, GR
中科院分区：
文献类型：
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作者：
Sathe, SS;Sizemore, N;Stark, GR
我们使用遗传方法生成了八种不同的突变人类细胞系，其中 NF-kappaB 被组成型激活。这些独立的克隆具有不同的表型，属于几个不同的遗传互补群体。在一个克隆中，kappaB (IkappaB) 激酶抑制剂具有组成型活性，但在其他七个克隆中则不然，尽管 IkappaB 在所有八个克隆中均被降解。因此，IkappaB 降解和 NF-kappaB 激活的 IkappaB 激酶独立机制在这些突变体中占主导地位。对突变体的生化分析表明，它们至少分为五个不同的类别，不同的上游激酶被激活，证实了 NF-kappaB 激活的多种机制。通过将具有组成型活性 NF-kappaB 的逆转录病毒 cDNA 文库引入 Ras C6 细胞系，然后进行功能互补的选择，我们分离了编码烯醇酶 1 C 末端片段的 cDNA，并将其鉴定为 NF-kappaB 的负调节因子。
We have used a genetic approach to generate eight different mutant human cell lines in which NF-kappaB is constitutively activated. These independent clones have different phenotypes and belong to several different genetic complementation groups. In one clone inhibitor of kappaB (IkappaB) kinase is constitutively active, but in the seven others it is not, despite the fact that IkappaB is degraded in all eight clones. Thus, IkappaB kinase-independent mechanisms of IkappaB degradation and NF-kappaB activation are predominant in these mutants. Biochemical analyses of the mutants revealed that they fall into at least five different categories, differing in the sets of upstream kinases that are activated, confirming multiple mechanisms of NF-kappaB activation. By introducing a retroviral cDNA library into the Ras C6 cell line, with constitutively active NF-kappaB, followed by selection for functional complementation, we isolated a cDNA encoding a C-terminal fragment of enolase 1 and identified it as negative regulator of NF-kappaB.