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兔网织红细胞双链RNA激活的真核起始因子3激酶的纯化及性质

Purification and properties of the double-stranded RNA-activated eukaryotic initiation factor 3 kinase from rabbit reticulocytes.

作者信息

Grosfeld H, Ochoa S

出版信息

Proc Natl Acad Sci U S A. 1980 Nov;77(11):6526-30. doi: 10.1073/pnas.77.11.6526.

DOI:10.1073/pnas.77.11.6526
PMID:6935666
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC350318/
Abstract

The double-stranded RNA (dsRNA)-activated protein kinase (DAI) that phosphorylates the alpha subunit of the eukaryotic initiation factor eIF-2 and inhibits chain initiation has been isolated from rabbit reticulocyte lysates. The nonactivated enzyme or the enzyme partially activated by incubation with low levels of dsRNA (pro-DAI) could be purified only to a slight extent. However, the enzyme that was fully activated by incubation with both dsRNA and ATP was purified to near homogeneity. Active DAI is a phosphoprotein with an apparent subunit mass of 68,000 daltons. It can phosphorylate histone as well as the alpha subunit of eIF-2. Our results suggest that, after interaction with dsRNA, the enzyme phosphorylates itself and is thereby activated to phosphorylate alpha eIF-2 and histone.

摘要

双链RNA（dsRNA）激活的蛋白激酶（DAI）可使真核起始因子eIF-2的α亚基磷酸化并抑制链起始，已从兔网织红细胞裂解物中分离出来。未激活的酶或通过与低水平dsRNA孵育而部分激活的酶（前DAI）仅能在一定程度上得到纯化。然而，通过与dsRNA和ATP一起孵育而完全激活的酶被纯化至接近均一性。活性DAI是一种磷蛋白，其亚基表观分子量为68,000道尔顿。它可以使组蛋白以及eIF-2的α亚基磷酸化。我们的结果表明，在与dsRNA相互作用后，该酶自身磷酸化，从而被激活以磷酸化αeIF-2和组蛋白。

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1
Purification and properties of the double-stranded RNA-activated eukaryotic initiation factor 3 kinase from rabbit reticulocytes.兔网织红细胞双链RNA激活的真核起始因子3激酶的纯化及性质
Proc Natl Acad Sci U S A. 1980 Nov;77(11):6526-30. doi: 10.1073/pnas.77.11.6526.
2
Characterization of double-stranded-RNA-activated kinase that phosphorylates alpha subunit of eukaryotic initiation factor 2 (eIF-2 alpha) in reticulocyte lysates.网织红细胞裂解物中使真核起始因子2（eIF-2α）的α亚基磷酸化的双链RNA激活激酶的特性分析
Proc Natl Acad Sci U S A. 1980 Feb;77(2):832-6. doi: 10.1073/pnas.77.2.832.
3
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Proc Natl Acad Sci U S A. 1980 Mar;77(3):1286-90. doi: 10.1073/pnas.77.3.1286.
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Double-stranded RNA-dependent phosphorylation of protein P1 and eukaryotic initiation factor 2 alpha does not correlate with protein synthesis inhibition in a cell-free system from interferon-treated mouse L cells.在经干扰素处理的小鼠L细胞的无细胞体系中，蛋白质P1和真核起始因子2α的双链RNA依赖性磷酸化与蛋白质合成抑制不相关。
Proc Natl Acad Sci U S A. 1983 Jan;80(1):41-5. doi: 10.1073/pnas.80.1.41.

引用本文的文献

1
Protein synthesis in rabbit reticulocytes: characteristics of the protein factor RF that reverses inhibition of protein synthesis in heme-deficient reticulocyte lysates.兔网织红细胞中的蛋白质合成：逆转血红素缺乏的网织红细胞裂解物中蛋白质合成抑制作用的蛋白质因子RF的特性。
Proc Natl Acad Sci U S A. 1982 Nov;79(21):6517-21. doi: 10.1073/pnas.79.21.6517.
2
Mode of action of the heme-controlled translational inhibitor: relationship of eukaryotic initiation factor 2-stimulating protein to translation restoring factor.血红素控制的翻译抑制剂的作用模式：真核起始因子2刺激蛋白与翻译恢复因子的关系。
Proc Natl Acad Sci U S A. 1981 Jan;78(1):220-3. doi: 10.1073/pnas.78.1.220.
3

本文引用的文献

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Protein measurement with the Folin phenol reagent.使用福林酚试剂进行蛋白质测定。
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Purification and characterization of a latent precursor of a double-stranded RNA dependent protein kinase from reticulocyte lysates.从网织红细胞裂解物中纯化和鉴定一种双链RNA依赖性蛋白激酶的潜在前体。
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Site-specific phosphorylation of the alpha subunit of eukaryotic initiation factor eIF-2 by the heme-regulated and double-stranded RNA-activated eIF-2 alpha kinases from rabbit reticulocyte lysates.
Regulation of double-stranded RNA-activated eukaryotic initiation factor 2 alpha kinase by type 2 protein phosphatase in reticulocyte lysates.
网织红细胞裂解物中2型蛋白磷酸酶对双链RNA激活的真核起始因子2α激酶的调节
Proc Natl Acad Sci U S A. 1982 Nov;79(21):6512-6. doi: 10.1073/pnas.79.21.6512.
4
Regulation of protein synthesis in vesicular stomatitis virus-infected mouse L-929 cells by decreased protein synthesis initiation factor 2 activity.通过降低蛋白质合成起始因子2的活性来调节水疱性口炎病毒感染的小鼠L-929细胞中的蛋白质合成
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Double-stranded RNA-dependent phosphorylation of protein P1 and eukaryotic initiation factor 2 alpha does not correlate with protein synthesis inhibition in a cell-free system from interferon-treated mouse L cells.在经干扰素处理的小鼠L细胞的无细胞体系中，蛋白质P1和真核起始因子2α的双链RNA依赖性磷酸化与蛋白质合成抑制不相关。
Proc Natl Acad Sci U S A. 1983 Jan;80(1):41-5. doi: 10.1073/pnas.80.1.41.
6
Presence of haemin-controlled eIF-2 alpha kinases in both undifferentiated and differentiating mouse erythroleukaemia cells.未分化和正在分化的小鼠红白血病细胞中均存在血红素控制的真核起始因子2α激酶。
Biochem J. 1989 Sep 1;262(2):569-74. doi: 10.1042/bj2620569.
7
Detection of activated double-stranded RNA-dependent protein kinase in 3T3-F442A cells.在3T3-F442A细胞中检测活化的双链RNA依赖性蛋白激酶。
Proc Natl Acad Sci U S A. 1988 Mar;85(5):1427-31. doi: 10.1073/pnas.85.5.1427.
兔网织红细胞裂解物中血红素调节的双链RNA激活的真核起始因子eIF-2的α亚基的位点特异性磷酸化。
Proc Natl Acad Sci U S A. 1980 Mar;77(3):1286-90. doi: 10.1073/pnas.77.3.1286.
4
Characterization of double-stranded-RNA-activated kinase that phosphorylates alpha subunit of eukaryotic initiation factor 2 (eIF-2 alpha) in reticulocyte lysates.网织红细胞裂解物中使真核起始因子2（eIF-2α）的α亚基磷酸化的双链RNA激活激酶的特性分析
Proc Natl Acad Sci U S A. 1980 Feb;77(2):832-6. doi: 10.1073/pnas.77.2.832.
5
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.在噬菌体T4头部组装过程中结构蛋白的切割
Nature. 1970 Aug 15;227(5259):680-5. doi: 10.1038/227680a0.
6
Control of globin synthesis by hemin: factors influencing formation of an inhibitor of globin chain initiation in reticulocyte lysates.血红素对珠蛋白合成的调控：影响网织红细胞裂解物中珠蛋白链起始抑制剂形成的因素。
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A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.一种利用蛋白质 - 染料结合原理对微克级蛋白质进行定量的快速灵敏方法。
Anal Biochem. 1976 May 7;72:248-54. doi: 10.1016/0003-2697(76)90527-3.
8
Additional evidence that the hemin-controlled translational repressor from rabbit reticulocytes is a protein kinase.来自兔网织红细胞的血红素控制的翻译阻遏物是一种蛋白激酶的更多证据。
Biochem Biophys Res Commun. 1977 Jan 24;74(2):559-69. doi: 10.1016/0006-291x(77)90340-0.
9
Analysis of phosphorylation of protein synthesis initiation factor eIF-2 by two-dimensional gel electrophoresis.通过二维凝胶电泳分析蛋白质合成起始因子eIF-2的磷酸化作用。
Eur J Biochem. 1978 Sep 1;89(2):517-21. doi: 10.1111/j.1432-1033.1978.tb12556.x.
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Inhibition of protein synthesis by double-stranded RNA and phosphorylation of initiation factor, eIF-2.双链RNA对蛋白质合成的抑制作用以及起始因子eIF-2的磷酸化
J Biol Chem. 1978 Jun 25;253(12):4219-23.