NF45 and NF90 Regulate HS4-dependent Interleukin-13 Transcription in T Cells
NF45 and NF90 Regulate HS4-dependent Interleukin-13 Transcription in T Cells
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DOI：
10.1074/jbc.m109.041004
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发表时间：
2010-03-12
影响因子：
4.8
通讯作者：
Vercelli, Donata
中科院分区：
文献类型：
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作者：
Kiesler, Patricia;Haynes, Paul A.;Vercelli, Donata
细胞因子白细胞介素-13（IL-13）的表达对于Th 2免疫应答和Th 2介导的过敏性疾病至关重要。人IL 13表达的激活涉及染色质重塑和在整个基因座中形成多个DNA酶I超敏感位点。其中，HS 4在初始和极化的CD 4（+）T细胞中的远端IL 13启动子中检测到。我们在本文中表明，HS 4在瞬时转染、激活的人CD 4（+）Jurkat T细胞和原代鼠Th 2细胞中充当IL 13近端启动子活性的位置独立性、方向依赖性正调节因子。通过DNA亲和层析结合串联质谱、染色质免疫沉淀和凝胶位移分析证实，HS 4的3 '-半（HS 4 - 3'）负责IL 13上调并结合核因子（NF）90和NF 45。值得注意的是，HS 4 -3'内的CTGTT NF 45/NF 90结合基序对于IL 13表达的HS 4依赖性上调是关键的。此外，将HS 4-IL 13报告载体转染到来自野生型、NF 45（+/-）或NF 90（+/-）小鼠的原代体外分化的Th 2细胞中显示，HS 4活性精确地依赖于内源性NF 45（+/-）的水平。（和较低程度的NF 90），因为在NF 45（+/-）细胞中HS 4依赖性IL 13表达实际上被消除，而在NF 90（+/-）细胞中减少。总的来说，我们的结果确定NF 45和NF 90作为响应T细胞活化的HS 4依赖性人IL 13转录的新调节因子。
Expression of the cytokine interleukin-13 (IL13) is critical for Th2 immune responses and Th2-mediated allergic diseases. Activation of human IL13 expression involves chromatin remodeling and formation of multiple DNase I-hypersensitive sites throughout the locus. Among these, HS4 is detected in the distal IL13 promoter in both naive and polarized CD4(+) T cells. We show herein that HS4 acts as a position-independent, orientation-dependent positive regulator of IL13 proximal promoter activity in transiently transfected, activated human CD4(+) Jurkat T cells and primary murine Th2 cells. The 3'-half of HS4 (HS4-3') was responsible for IL13 up-regulation and bound nuclear factor (NF) 90 and NF45, as demonstrated by DNA affinity chromatography coupled with tandem mass spectrometry, chromatin immunoprecipitation, and gel shift analysis. Notably, the CTGTT NF45/NF90-binding motif within HS4-3' was critical for HS4-dependent upregulation of IL13 expression. Moreover, transfection of HS4-IL13 reporter vectors into primary, in vitro differentiated Th2 cells from wild-type, NF45(+/-), or NF90(+/-) mice showed that HS4 activity was exquisitely dependent on the levels of endogenous NF45 (and to a lesser degree NF90), because HS4-dependent IL13 expression was virtually abrogated in NF45(+/-) cells and reduced in NF90(+/-) cells. Collectively, our results identify NF45 and NF90 as novel regulators of HS4-dependent human IL13 transcription in response to T cell activation.