Inducible expression of IκBα repressor mutants interferes with NF-κB activity and HIV-1 replication in Jurkat T cells
Inducible expression of IκBα repressor mutants interferes with NF-κB activity and HIV-1 replication in Jurkat T cells
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DOI：
10.1074/jbc.273.13.7431
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发表时间：
1998-03-27
影响因子：
4.8
通讯作者：
Hiscott, J
中科院分区：
文献类型：
--
作者：
Kwon, H;Pelletier, N;Hiscott, J
人类免疫缺陷病毒（HIV-1）利用NF-κ B/Rel蛋白通过HIV-1长末端重复序列（LTR）中的NF-κ B结合位点来调节转录。正常情况下，NF-κ B通过抑制性I κ.B蛋白保留在细胞质中;在包括病毒在内的多种激活剂刺激后，I κ B α被磷酸化并降解，导致NF-κ B释放。在本研究中，我们使用稳定选择的Jurkat T细胞检测了四环素诱导的I κ B α（TD-I κ B α）的反式显性阻遏物表达对HIV-1增殖的影响，TD-I κ B α早在加入多西环素后3 h就可诱导表达，并显著降低NF-κ B DNA结合活性和LTR指导的基因活性。有趣的是，诱导的TD-I κ B α表达也在诱导后24 h将内源性I κ B α表达降低至不可检测的水平，表明TD-I κ B α抑制内源性NF-κ B依赖性基因转录。TD-I κ B α表达也使Jurkat细胞对肿瘤坏死因子诱导的凋亡敏感。通过p24抗原、逆转录酶和病毒RNA测量，TD-1 kappa B alpha诱导显着改变了Jurbat细胞的从头HIV-1感染，从而抑制了HIV-1增殖。考虑到NF-κ B/I κ B通路的多种功能，TD-I κ B α表达可能在几个水平上干扰HIV-1增殖：LTR介导的转录、Rev介导的病毒RNA输出、HIV-1诱导的促炎细胞因子的抑制以及HIV-1感染细胞对凋亡的敏感性增加。
Human immunodeficiency virus (HIV-1) utilizes the NF-kappa B/Rel proteins to regulate transcription through NF-kappa B binding sites in the HIV-1 long terminal repeat (LTR). Normally, NF-kappa B is retained in the cytoplasm by inhibitory I kappa.B proteins; after stimulation by multiple activators including viruses, I kappa B alpha is phosphorylated and degraded, resulting in NF-kappa B release. In the present study, we examined the effect of tetracycline inducible expression of transdominant repressors of I kappa B alpha (TD-I kappa B alpha) on HIV-1 multiplication using stably selected Jurkat T cells, TD-I kappa B alpha was inducibly expressed as early as 3 h after doxycycline addition and dramatically reduced both NF-kappa B DNA binding activity and LTR directed gene activity. Interestingly, induced TD-I kappa B alpha expression also decreased endogenous I kappa B alpha expression to undetectable levels by 24 h after induction, demonstrating that TD-I kappa B alpha repressed endogenous NF-kappa B-dependent gene transcription. TD-I kappa B alpha expression also sensitized Jurkat cells to tumor necrosis factor-induced apoptosis. De novo HIV-1 infection of Jurbat cells was dramatically altered by TD-I kappa B alpha induction, resulting in inhibition of HIV-1 multiplication, as measured by p24 antigen, reverse transcriptase, and viral RNA. Given the multiple functions of the NF-kappa B/I kappa B pathway, TD-I kappa B alpha expression may interfere with HIV-1 multiplication at several levels: LTR-mediated transcription, Rev-mediated export of viral RNA, inhibition of HIV-1-induced pro-inflammatory cytokines, and increased sensitivity of HIV-1-infected cells to apoptosis.