Gupta Manish, Rath Pramod C
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.
Int J Biol Macromol. 2014 Apr;65:41-50. doi: 10.1016/j.ijbiomac.2014.01.005. Epub 2014 Jan 9.
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.
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”。