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干扰素作用:由基因200簇中的Ifi 200基因编码的干扰素诱导型52-kD蛋白的细胞质和细胞核定位。

Interferon action: cytoplasmic and nuclear localization of the interferon-inducible 52-kD protein that is encoded by the Ifi 200 gene from the gene 200 cluster.

作者信息

Choubey D, Lengyel P

机构信息

Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511.

出版信息

J Interferon Res. 1993 Feb;13(1):43-52. doi: 10.1089/jir.1993.13.43.

Abstract

Recently, we reported that an interferon (IFN)-inducible, murine 72-kD phosphoprotein (the 204 protein) that is encoded by the Ifi 204 gene from the gene 200 cluster is localized in the nucleolus and the nucleoplasm. We have now raised a polyclonal antiserum against the 202 protein that is encoded by the Ifi 202 gene from the same gene cluster and regions of which are homologous to those from the 204 protein. Using the antiserum, we established that the 202 protein is a 52-kD phosphoprotein whose level in cells from various murine lines can be increased up to 16-fold upon treatment with IFN-alpha. Experiments involving fractionation of cell lysates and indirect immunofluorescence microscopy of cultured cells revealed that the 202 protein was localized in the cytoplasm and the nucleus. Upon treatment of cells with IFN, the 202 protein first accumulated on the surface of a cytoplasmic, membranous fraction and after prolonged treatment with IFN it was localized mainly in the nucleus. In IFN-treated mitotic AKR cells, the 202 protein was colocalized with chromosomes. 202 protein extracted from IFN-treated AKR cells bound double-stranded DNA in vitro. Studies on 202 protein function should be facilitated by the availability of complete cDNA clones and the finding of cell lines and an inbred strain of mice in which the expression of this protein was impaired.

摘要

最近,我们报道了一种由基因200簇中的Ifi 204基因编码的干扰素(IFN)诱导型小鼠72-kD磷蛋白(204蛋白)定位于核仁和核质中。我们现在制备了一种针对由同一基因簇中的Ifi 202基因编码的202蛋白的多克隆抗血清,该蛋白的区域与204蛋白的区域同源。使用该抗血清,我们确定202蛋白是一种52-kD磷蛋白,在用IFN-α处理后,各种小鼠品系细胞中的该蛋白水平可增加至16倍。涉及细胞裂解物分级分离和培养细胞间接免疫荧光显微镜检查的实验表明,202蛋白定位于细胞质和细胞核中。在用IFN处理细胞后,202蛋白首先积聚在细胞质膜性部分的表面,在用IFN长时间处理后,它主要定位于细胞核中。在经IFN处理的有丝分裂AKR细胞中,202蛋白与染色体共定位。从经IFN处理的AKR细胞中提取的202蛋白在体外与双链DNA结合。完整cDNA克隆的可用性以及发现该蛋白表达受损的细胞系和近交系小鼠将有助于对202蛋白功能的研究。

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