Sweet M J, Stacey K J, Ross I L, Ostrowski M C, Hume D A
Department of Microbiology, University of Queensland, Brisbane, Australia.
J Inflamm. 1998;48(2):67-83.
The HIV-1 promoter was used as a model to identify transcription factors involved in LPS-dependent transcription in RAW 264 murine macrophages. Expression plasmids for Ets-2 and PU.1 trans-activated the HIV-1 LTR and recombinant PU.1 and an Ets-2 DNA binding domain/GST fusion protein bound to the 5' kappa B site of the HIV-1 enhancer. Ets-2 mRNA was LPS-inducible in RAW 264 cells and LPS stimulated phosphorylation of threonine 72 residue within the Ets-2 pointed domain. Induction of Ets-2 and other LPS-responsive transcription factors was also observed upon addition of plasmid DNA, which complicates interpretation of transient transfections. The proximal promoter region, containing two Sp1 sites, was also LPS-responsive. We propose that the kappa B elements and the tandem Sp1 sites act as LPS response elements and that kappa B-mediated LPS action involves Ets and rel factors.
HIV-1启动子被用作模型，以鉴定参与RAW 264小鼠巨噬细胞中脂多糖（LPS）依赖性转录的转录因子。Ets-2和PU.1的表达质粒可反式激活HIV-1长末端重复序列（LTR），重组PU.1和Ets-2 DNA结合结构域/谷胱甘肽S-转移酶（GST）融合蛋白可与HIV-1增强子的5' κB位点结合。Ets-2 mRNA在RAW 264细胞中可被LPS诱导，且LPS刺激Ets-2尖形结构域内苏氨酸72残基的磷酸化。加入质粒DNA后也观察到Ets-2和其他LPS反应性转录因子的诱导，这使得瞬时转染的解释变得复杂。包含两个Sp1位点的近端启动子区域也对LPS有反应。我们提出κB元件和串联Sp1位点作为LPS反应元件，且κB介导的LPS作用涉及Ets和rel因子。
