A cell-based bioluminescence assay reveals dose-dependent and contextual repression of AP-1-driven gene expression by BACH2.
A cell-based bioluminescence assay reveals dose-dependent and contextual repression of AP-1-driven gene expression by BACH2.
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DOI：
10.1038/s41598-020-75732-z
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发表时间：
2020-11-03
影响因子：
4.6
通讯作者：
Roychoudhuri R
中科院分区：
文献类型：
--
作者：
Vardaka P;Lozano T;Bot C;Ellery J;Whiteside SK;Imianowski CJ;Farrow S;Walker S;Okkenhaug H;Yang J;Okkenhaug K;Kuo P;Roychoudhuri R
尽管效应CD 4+和CD 8 + T细胞促进免疫活化并可驱动感染和癌症的清除，但CD 4+调节性T（Treg）细胞抑制其功能，有助于免疫稳态和癌症免疫抑制。转录因子BACH 2作为T细胞分化的普遍调节剂发挥作用，促进CD 4 + Treg细胞的发育并抑制多效应T细胞（Teff）谱系的效应子功能。在这里，我们报告了一个稳定的基于细胞的生物发光测定BACH 2的转录因子活性的发展。四环素诱导的BACH 2表达导致抑制佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）/离子霉素驱动的荧光素酶报告基因的激活，该报告基因含有来自小鼠Ifng +18 k增强子的BACH 2/AP-1靶序列。BACH 2表达以剂量依赖性方式抑制荧光素酶信号，但在高水平AP-1信号传导时这种活性被消除，表明BACH 2对AP-1驱动的基因表达的背景调节。最后，使用开发的报告分析，我们发现，组蛋白脱乙酰酶3（HDAC 3）的选择性抑制剂，RGFP 966，抑制BACH 2介导的信号驱动的荧光素酶表达的抑制。除了能够进行机制研究之外，这种基于细胞的报告基因还可以鉴定BACH 2功能的小分子激动剂或拮抗剂，用于药物开发。
Whereas effector CD4+ and CD8+ T cells promote immune activation and can drive clearance of infections and cancer, CD4+ regulatory T (Treg) cells suppress their function, contributing to both immune homeostasis and cancer immunosuppression. The transcription factor BACH2 functions as a pervasive regulator of T cell differentiation, promoting development of CD4+ Treg cells and suppressing the effector functions of multiple effector T cell (Teff) lineages. Here, we report the development of a stable cell-based bioluminescence assay of the transcription factor activity of BACH2. Tetracycline-inducible BACH2 expression resulted in suppression of phorbol 12-myristate 13-acetate (PMA)/ionomycin-driven activation of a luciferase reporter containing BACH2/AP-1 target sequences from the mouse Ifng + 18k enhancer. BACH2 expression repressed the luciferase signal in a dose-dependent manner but this activity was abolished at high levels of AP-1 signalling, suggesting contextual regulation of AP-1 driven gene expression by BACH2. Finally, using the reporter assay developed, we find that the histone deacetylase 3 (HDAC3)-selective inhibitor, RGFP966, inhibits BACH2-mediated repression of signal-driven luciferase expression. In addition to enabling mechanistic studies, this cell-based reporter may enable identification of small molecule agonists or antagonists of BACH2 function for drug development.
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影响因子：
4.4
作者：
Luisier, Florian;Vonesch, Cedric;Unser, Michael
通讯作者：
Unser, Michael
DOI：
10.1126/science.1233465
发表时间：
2013-05-10
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Reinke AW;Baek J;Ashenberg O;Keating AE
通讯作者：
Keating AE
影响因子：
56.9
作者：
TURNER, R;TJIAN, R
通讯作者：
TJIAN, R
影响因子：
16.6
作者：
Kuwahara, Makoto;Ise, Wataru;Ochi, Mizuki;Suzuki, Junpei;Kometani, Kohei;Maruyama, Saho;Izumoto, Maya;Matsumoto, Akira;Takemori, Nobuaki;Takemori, Ayako;Shinoda, Kenta;Nakayama, Toshinori;Ohara, Osamu;Yasukawa, Masaki;Sawasaki, Tatsuya;Kurosaki, Tomohiro;Yamashita, Masakatsu
通讯作者：
Yamashita, Masakatsu
影响因子：
29.7
作者：
Josefowicz SZ;Lu LF;Rudensky AY
通讯作者：
Rudensky AY