Induction of the human protein P56 by interferon, double-stranded RNA, or virus infection
Induction of the human protein P56 by interferon, double-stranded RNA, or virus infection
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DOI：
10.1006/viro.1999.0135
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发表时间：
2000-02-15
期刊：
影响因子：
3.7
通讯作者：
Sen, GC
中科院分区：
文献类型：
--
作者：
Guo, JJ;Peters, KL;Sen, GC
P56是干扰素（IFN）处理人类细胞诱导的最丰富的蛋白。为了便于对其诱导模式和细胞功能的研究，我们在大肠杆菌中以六组氨酸标记的蛋白形式表达重组P56，并采用亲和层析纯化，使其具有明显的同质性。针对该重组蛋白的多克隆抗体表明P56主要是一个细胞质蛋白。转染P56的细胞表达对水疱性口炎病毒和脑心肌炎病毒的复制没有抑制作用。ifn - β能快速诱导P56的合成，蛋白的半衰期为6 h。ifn - γ或poly(a)（+）不能诱导P56的合成，但poly(I)-poly(C)或85 bp的合成双链RNA能有效诱导P56的合成。同样，当缺乏I型IFN基因的GRE细胞被水疱性口炎病毒、脑心肌炎病毒或仙台病毒感染时，可诱导P56。令人惊讶的是，仙台病毒也可以在突变细胞系P2.1中诱导P56，该细胞系对ifn - α / β或双链RNA都没有反应。病毒感染P2.1细胞和亲本U4C细胞诱导P56之前，IRF-3被激活，从细胞质向细胞核的易位判断。(C) 2000年学术出版社。
P56 is the most abundant protein induced by interferon (IFN) treatment of human cells. To facilitate studies on its induction pattern and cellular functions, we expressed recombinant P56 as a hexahistidine-tagged protein in Escherichia coli and purified it to apparent homogeneity using affinity chromatography. A polyclonal antibody raised against this recombinant protein was used to show that P56 is primarily a cytoplasmic protein. Cellular expression of P56 by transfection did not inhibit the replication of vesicular stomatitis virus and encephalomyocarditis virus. P56 synthesis was rapidly induced by IFN-beta, and the protein had a half-life of 6 h. IFN-gamma or poly(A)(+) could not induce the protein, but poly(I)-poly(C) or an 85-bp synthetic double-stranded RNA efficiently induced it. Similarly, infection of GRE cells, which are devoid of type I IFN genes, by vesicular stomatitis virus, encephalomyocarditis virus, or Sendai virus caused P56 induction. Surprisingly, Sendai virus could also induce P56 in the mutant cell line P2.1, which cannot respond to either IFN-alpha/beta or double-stranded RNA. Induction of P56 in the P2.1 cells and the parental U4C cells by virus infection was preceded by activation of IRF-3 as judged by its translocation to the nucleus from the cytoplasm. (C) 2000 Academic Press.