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重组杆状病毒表达的铁调节因子：RNA结合脱辅基蛋白与含Fe-S簇的乌头酸酶之间的转化

Iron regulatory factor expressed from recombinant baculovirus: conversion between the RNA-binding apoprotein and Fe-S cluster containing aconitase.

作者信息

Emery-Goodman A, Hirling H, Scarpellino L, Henderson B, Kühn L C

机构信息

Swiss Institute for Experimental Cancer Research, Genetics Unit, Epalinges.

出版信息

Nucleic Acids Res. 1993 Mar 25;21(6):1457-61. doi: 10.1093/nar/21.6.1457.

DOI:10.1093/nar/21.6.1457
PMID:8464737
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC309332/
Abstract

Iron regulatory factor (IRF) is a cytoplasmic mRNA-binding protein that coordinates post-transcriptionally the expression of several important proteins in iron metabolism. Binding of IRF to iron-responsive elements (IRE) in the 5' untranslated region (UTR) of ferritin and erythroid 5-aminolevulinic acid-synthase mRNAs inhibits their translation, whereas binding to IREs in the 3' UTR of transferrin receptor (TfR) mRNA prevents the degradation of this mRNA. IRF binds RNA strongly after iron deprivation, but is inactive, yet present, under conditions of high cellular iron supply. Recently, IRF was also shown to have aconitase activity indicating the existence of an Fe-S cluster in the protein. In the current study we expressed human IRF in insect cells from recombinant baculovirus and analysed IRE-binding and aconitase activities under various culture conditions. Newly made apoprotein, synthesized in the absence of iron, was fully active in IRE-binding, but showed no aconitase activity. In contrast, IRF made by cells grown in high iron medium bound RNA poorly, but exhibited high aconitase activity with a Km of 9.2 microM for cis-aconitate. Apo-IRF was converted in vitro to active aconitase by Fe-S cluster-generating conditions, and under the same conditions lost its RNA-binding capacity. These results indicate that the two activities are mutually exclusive and controlled through formation of the Fe-S cluster.

摘要

铁调节因子（IRF）是一种细胞质mRNA结合蛋白，它在转录后协调铁代谢中几种重要蛋白质的表达。IRF与铁蛋白和红细胞5-氨基酮戊酸合酶mRNA的5'非翻译区（UTR）中的铁反应元件（IRE）结合会抑制它们的翻译，而与转铁蛋白受体（TfR）mRNA的3'UTR中的IRE结合则可防止该mRNA的降解。铁缺乏后，IRF会强烈结合RNA，但在细胞铁供应充足的情况下，它虽存在却无活性。最近，IRF还被证明具有乌头酸酶活性，这表明该蛋白中存在一个铁硫簇。在本研究中，我们利用重组杆状病毒在昆虫细胞中表达了人IRF，并分析了其在各种培养条件下的IRE结合活性和乌头酸酶活性。在无铁条件下合成的新生成的脱辅基蛋白在IRE结合方面完全有活性，但没有乌头酸酶活性。相反，在高铁培养基中生长的细胞产生的IRF与RNA的结合能力较差，但对顺乌头酸表现出高乌头酸酶活性，其Km为9.2微摩尔。脱辅基IRF在体外通过产生铁硫簇的条件转化为有活性的乌头酸酶，并且在相同条件下失去了其RNA结合能力。这些结果表明这两种活性相互排斥，并通过铁硫簇的形成来控制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7046/309332/e3e4a826b3f1/nar00055-0126-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7046/309332/e3e4a826b3f1/nar00055-0126-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7046/309332/e3e4a826b3f1/nar00055-0126-a.jpg

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本文引用的文献

1
The role of iron in the activation-inactivation of aconitase.铁在乌头酸酶激活-失活过程中的作用。
J Biol Chem. 1983 Sep 25;258(18):11098-105.
2
Purification and kinetic studies of beef liver cytoplasmic aconitase.牛肝细胞质乌头酸酶的纯化及动力学研究
J Biol Chem. 1967 Sep 10;242(17):3833-8.
3
Modification and secretion of human interleukin 2 produced in insect cells by a baculovirus expression vector.杆状病毒表达载体在昆虫细胞中产生的人白细胞介素2的修饰与分泌
慢生根瘤菌的乌头酸酶基因（acnA）对其自由生活生长很重要，但对有效的根瘤共生并不重要。
J Bacteriol. 1996 Nov;178(21):6166-72. doi: 10.1128/jb.178.21.6166-6172.1996.
4
Molecular control of vertebrate iron metabolism: mRNA-based regulatory circuits operated by iron, nitric oxide, and oxidative stress.脊椎动物铁代谢的分子调控：由铁、一氧化氮和氧化应激操纵的基于mRNA的调控回路。
Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8175-82. doi: 10.1073/pnas.93.16.8175.
5
Nitric oxide and oxidative stress (H2O2) control mammalian iron metabolism by different pathways.一氧化氮和氧化应激（过氧化氢）通过不同途径调控哺乳动物的铁代谢。
Mol Cell Biol. 1996 Jul;16(7):3781-8. doi: 10.1128/MCB.16.7.3781.
6
The iron-responsive element-binding protein: localization of the RNA-binding site to the aconitase active-site cleft.铁反应元件结合蛋白：RNA结合位点定位于乌头酸酶活性位点裂隙处。
Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):574-8. doi: 10.1073/pnas.91.2.574.
7
Iron regulatory protein prevents binding of the 43S translation pre-initiation complex to ferritin and eALAS mRNAs.铁调节蛋白可阻止43S翻译起始前复合物与铁蛋白和eALAS mRNA结合。
EMBO J. 1994 Aug 15;13(16):3882-91. doi: 10.1002/j.1460-2075.1994.tb06699.x.
8
The bifunctional iron-responsive element binding protein/cytosolic aconitase: the role of active-site residues in ligand binding and regulation.双功能铁反应元件结合蛋白/胞质乌头酸酶：活性位点残基在配体结合和调节中的作用
Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7321-5. doi: 10.1073/pnas.91.15.7321.
9
Translational enhancement of H-ferritin mRNA by interleukin-1 beta acts through 5' leader sequences distinct from the iron responsive element.白细胞介素-1β对H-铁蛋白mRNA的翻译增强作用通过不同于铁反应元件的5'前导序列发挥作用。
Nucleic Acids Res. 1994 Jul 11;22(13):2678-86. doi: 10.1093/nar/22.13.2678.
10
Rapid responses to oxidative stress mediated by iron regulatory protein.铁调节蛋白介导的对氧化应激的快速反应。
EMBO J. 1995 Jun 15;14(12):2917-24. doi: 10.1002/j.1460-2075.1995.tb07291.x.
Proc Natl Acad Sci U S A. 1985 Dec;82(24):8404-8. doi: 10.1073/pnas.82.24.8404.
4
Murine cytotoxic activated macrophages inhibit aconitase in tumor cells. Inhibition involves the iron-sulfur prosthetic group and is reversible.小鼠细胞毒性活化巨噬细胞可抑制肿瘤细胞中的顺乌头酸酶。这种抑制作用涉及铁硫辅基，且是可逆的。
J Clin Invest. 1986 Sep;78(3):790-7. doi: 10.1172/JCI112642.
5
Noncoding 3' sequences of the transferrin receptor gene are required for mRNA regulation by iron.转铁蛋白受体基因的非编码3'序列是铁对mRNA进行调控所必需的。
EMBO J. 1987 May;6(5):1287-93. doi: 10.1002/j.1460-2075.1987.tb02366.x.
6
Binding of a cytosolic protein to the iron-responsive element of human ferritin messenger RNA.一种胞质蛋白与人铁蛋白信使核糖核酸的铁反应元件的结合。
Science. 1988 Sep 2;241(4870):1207-10. doi: 10.1126/science.3413484.
7
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8
Cytoplasmic protein binds in vitro to a highly conserved sequence in the 5' untranslated region of ferritin heavy- and light-subunit mRNAs.细胞质蛋白在体外与铁蛋白重链和轻链亚基mRNA的5'非翻译区中的一个高度保守序列结合。
Proc Natl Acad Sci U S A. 1988 Apr;85(7):2171-5. doi: 10.1073/pnas.85.7.2171.
9
Primary structure of the human 4F2 antigen heavy chain predicts a transmembrane protein with a cytoplasmic NH2 terminus.人类4F2抗原重链的一级结构预测其为一种跨膜蛋白，氨基末端位于细胞质内。
J Biol Chem. 1987 Jul 15;262(20):9574-80.
10
The state of cluster SH and S2- of aconitase during cluster interconversions and removal. A convenient preparation of apoenzyme.簇间相互转化和去除过程中乌头酸酶的簇SH和S2-状态。脱辅基酶的简便制备方法。
J Biol Chem. 1988 Jun 15;263(17):8194-8.