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干扰素调节因子-1(IRF-1)在小鼠骨髓细胞的细胞质中与发育和DNA损伤反应调节因子2(REDD2)相互作用。

Interferon regulatory factor-1 (IRF-1) interacts with regulated in development and DNA damage response 2 (REDD2) in the cytoplasm of mouse bone marrow cells.

作者信息

Gupta Manish, Rath Pramod C

机构信息

Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India.

Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India.

出版信息

Int J Biol Macromol. 2014 Apr;65:41-50. doi: 10.1016/j.ijbiomac.2014.01.005. Epub 2014 Jan 9.

Abstract

IRF-1 is a critical hematopoietic transcription factor, which regulates cell growth, development of immune cells, immune response, tumor suppression, apoptosis and autophagy in mammalian cells. Protein-protein interactions of IRF-1 in mouse bone marrow cells (BMCs) by GST-IRF-1 pull-down followed by mass spectrometry, coimmunoprecipitation, immunoblotting and colocalization show that regulated in development and DNA damage response 2 (REDD2) is an IRF-1-interacting protein. REDD2 is a highly conserved mammalian regulatory protein of the TSC2/mTOR pathway. It is structurally similar to REDD1 but has a distinct loop region. Cellular IRF-1 and REDD2 complex is present in the cytoplasm of BMCs as distinct speckles in punctate pattern. In vitro interaction of recombinant IRF-1 and REDD2 shows their physical interaction. Taken together, our results suggest that IRF-1 physically interacts with REDD2 in the large cytoplasmic protein complex, which may function as cellular signaling proteins for 'cross-talk' of mTOR and cytokine pathways during regulation of cell growth/proliferation, apoptosis and autophagy of mammalian bone marrow cells during health and disease.

摘要

干扰素调节因子1(IRF-1)是一种关键的造血转录因子,它在哺乳动物细胞中调节细胞生长、免疫细胞发育、免疫反应、肿瘤抑制、细胞凋亡和自噬。通过谷胱甘肽S-转移酶(GST)-IRF-1下拉法,随后进行质谱分析、免疫共沉淀、免疫印迹和共定位,对小鼠骨髓细胞(BMC)中IRF-1的蛋白质-蛋白质相互作用进行研究,结果表明,发育和DNA损伤反应调节因子2(REDD2)是一种与IRF-1相互作用的蛋白质。REDD2是TSC2/mTOR途径中一种高度保守的哺乳动物调节蛋白。它在结构上与REDD1相似,但有一个独特的环区。细胞内的IRF-1和REDD2复合物以点状模式存在于BMC细胞质中的不同斑点中。重组IRF-1和REDD2的体外相互作用表明了它们之间的物理相互作用。综上所述,我们的结果表明,IRF-1在大的细胞质蛋白复合物中与REDD2发生物理相互作用,这可能作为细胞信号蛋白,在健康和疾病状态下调节哺乳动物骨髓细胞的生长/增殖、细胞凋亡和自噬过程中,参与mTOR和细胞因子途径的“串扰”。

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