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1
Modulation of tRNA(iMet), eIF-2, and eIF-2B expression shows that GCN4 translation is inversely coupled to the level of eIF-2.GTP.Met-tRNA(iMet) ternary complexes.对起始甲硫氨酸转运核糖核酸（tRNA<sup>(iMet)</sup>）、真核起始因子2（eIF-2）和真核起始因子2B（eIF-2B）表达的调节表明，GCN4的翻译与eIF-2·GTP·甲硫氨酰-tRNA<sup>(iMet)</sup>三元复合物的水平呈负相关。
Mol Cell Biol. 1995 Nov;15(11):6351-63. doi: 10.1128/MCB.15.11.6351.
2
A protein complex of translational regulators of GCN4 mRNA is the guanine nucleotide-exchange factor for translation initiation factor 2 in yeast.GCN4 mRNA翻译调控因子的一种蛋白质复合物是酵母中翻译起始因子2的鸟嘌呤核苷酸交换因子。
Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5350-4. doi: 10.1073/pnas.90.11.5350.
3
Requirements for intercistronic distance and level of eukaryotic initiation factor 2 activity in reinitiation on GCN4 mRNA vary with the downstream cistron.在GCN4信使核糖核酸（mRNA）上重新起始时，顺反子间距离的要求以及真核起始因子2活性水平会因下游顺反子而异。
Mol Cell Biol. 1994 Apr;14(4):2616-28. doi: 10.1128/mcb.14.4.2616-2628.1994.
4
Gene-specific translational control of the yeast GCN4 gene by phosphorylation of eukaryotic initiation factor 2.通过真核起始因子2的磷酸化对酵母GCN4基因进行基因特异性翻译调控。
Mol Microbiol. 1993 Oct;10(2):215-23. doi: 10.1111/j.1365-2958.1993.tb01947.x.
5
Mutations in the GCD7 subunit of yeast guanine nucleotide exchange factor eIF-2B overcome the inhibitory effects of phosphorylated eIF-2 on translation initiation.酵母鸟嘌呤核苷酸交换因子eIF-2B的GCD7亚基中的突变克服了磷酸化eIF-2对翻译起始的抑制作用。
Mol Cell Biol. 1994 May;14(5):3208-22. doi: 10.1128/mcb.14.5.3208-3222.1994.
6
GCD10, a translational repressor of GCN4, is the RNA-binding subunit of eukaryotic translation initiation factor-3.GCD10是GCN4的一种翻译阻遏物，是真核生物翻译起始因子3的RNA结合亚基。
Genes Dev. 1995 Jul 15;9(14):1781-96. doi: 10.1101/gad.9.14.1781.
7
Evidence that GCD6 and GCD7, translational regulators of GCN4, are subunits of the guanine nucleotide exchange factor for eIF-2 in Saccharomyces cerevisiae.有证据表明，GCN4的翻译调节因子GCD6和GCD7是酿酒酵母中eIF-2的鸟嘌呤核苷酸交换因子的亚基。
Mol Cell Biol. 1993 Mar;13(3):1920-32. doi: 10.1128/mcb.13.3.1920-1932.1993.
8
Mammalian eukaryotic initiation factor 2 alpha kinases functionally substitute for GCN2 protein kinase in the GCN4 translational control mechanism of yeast.哺乳动物真核生物起始因子2α激酶在酵母的GCN4翻译控制机制中功能上替代了GCN2蛋白激酶。
Proc Natl Acad Sci U S A. 1993 May 15;90(10):4616-20. doi: 10.1073/pnas.90.10.4616.
9
Multicopy tRNA genes functionally suppress mutations in yeast eIF-2 alpha kinase GCN2: evidence for separate pathways coupling GCN4 expression to unchanged tRNA.多拷贝tRNA基因在功能上抑制酵母eIF-2α激酶GCN2中的突变：将GCN4表达与未改变的tRNA偶联的独立途径的证据。
Mol Cell Biol. 1994 Dec;14(12):7920-32. doi: 10.1128/mcb.14.12.7920-7932.1994.
10
GCD2, a translational repressor of the GCN4 gene, has a general function in the initiation of protein synthesis in Saccharomyces cerevisiae.GCD2是GCN4基因的一种翻译阻遏物，在酿酒酵母的蛋白质合成起始过程中具有一般性功能。
Mol Cell Biol. 1991 Jun;11(6):3203-16. doi: 10.1128/mcb.11.6.3203-3216.1991.

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本文引用的文献

1
A protein complex of translational regulators of GCN4 mRNA is the guanine nucleotide-exchange factor for translation initiation factor 2 in yeast.GCN4 mRNA翻译调控因子的一种蛋白质复合物是酵母中翻译起始因子2的鸟嘌呤核苷酸交换因子。
Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5350-4. doi: 10.1073/pnas.90.11.5350.
2
Evidence that GCD6 and GCD7, translational regulators of GCN4, are subunits of the guanine nucleotide exchange factor for eIF-2 in Saccharomyces cerevisiae.有证据表明，GCN4的翻译调节因子GCD6和GCD7是酿酒酵母中eIF-2的鸟嘌呤核苷酸交换因子的亚基。
Mol Cell Biol. 1993 Mar;13(3):1920-32. doi: 10.1128/mcb.13.3.1920-1932.1993.
3
GCD11, a negative regulator of GCN4 expression, encodes the gamma subunit of eIF-2 in Saccharomyces cerevisiae.GCD11是GCN4表达的负调控因子，在酿酒酵母中编码eIF-2的γ亚基。
Mol Cell Biol. 1993 Jan;13(1):506-20. doi: 10.1128/mcb.13.1.506-520.1993.
4
Translation of the yeast transcriptional activator GCN4 is stimulated by purine limitation: implications for activation of the protein kinase GCN2.酵母转录激活因子GCN4的翻译受嘌呤限制的刺激：对蛋白激酶GCN2激活的影响。
Mol Cell Biol. 1993 Aug;13(8):5099-111. doi: 10.1128/mcb.13.8.5099-5111.1993.
5
Guanine nucleotide exchange factor for eukaryotic translation initiation factor 2 in Saccharomyces cerevisiae: interactions between the essential subunits GCD2, GCD6, and GCD7 and the regulatory subunit GCN3.酿酒酵母中真核生物翻译起始因子2的鸟嘌呤核苷酸交换因子：必需亚基GCD2、GCD6和GCD7与调节亚基GCN3之间的相互作用
Mol Cell Biol. 1993 Aug;13(8):4618-31. doi: 10.1128/mcb.13.8.4618-4631.1993.
6
Mutations in the GCD7 subunit of yeast guanine nucleotide exchange factor eIF-2B overcome the inhibitory effects of phosphorylated eIF-2 on translation initiation.酵母鸟嘌呤核苷酸交换因子eIF-2B的GCD7亚基中的突变克服了磷酸化eIF-2对翻译起始的抑制作用。
Mol Cell Biol. 1994 May;14(5):3208-22. doi: 10.1128/mcb.14.5.3208-3222.1994.
7
Mutations in the alpha subunit of eukaryotic translation initiation factor 2 (eIF-2 alpha) that overcome the inhibitory effect of eIF-2 alpha phosphorylation on translation initiation.真核生物翻译起始因子2（eIF-2α）α亚基中的突变可克服eIF-2α磷酸化对翻译起始的抑制作用。
Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7215-9. doi: 10.1073/pnas.90.15.7215.
8
Mammalian eukaryotic initiation factor 2 alpha kinases functionally substitute for GCN2 protein kinase in the GCN4 translational control mechanism of yeast.哺乳动物真核生物起始因子2α激酶在酵母的GCN4翻译控制机制中功能上替代了GCN2蛋白激酶。
Proc Natl Acad Sci U S A. 1993 May 15;90(10):4616-20. doi: 10.1073/pnas.90.10.4616.
9
Multicopy tRNA genes functionally suppress mutations in yeast eIF-2 alpha kinase GCN2: evidence for separate pathways coupling GCN4 expression to unchanged tRNA.多拷贝tRNA基因在功能上抑制酵母eIF-2α激酶GCN2中的突变：将GCN4表达与未改变的tRNA偶联的独立途径的证据。
Mol Cell Biol. 1994 Dec;14(12):7920-32. doi: 10.1128/mcb.14.12.7920-7932.1994.
10
Gene-specific translational control of the yeast GCN4 gene by phosphorylation of eukaryotic initiation factor 2.通过真核起始因子2的磷酸化对酵母GCN4基因进行基因特异性翻译调控。
Mol Microbiol. 1993 Oct;10(2):215-23. doi: 10.1111/j.1365-2958.1993.tb01947.x.

对起始甲硫氨酸转运核糖核酸（tRNA<sup>(iMet)</sup>）、真核起始因子2（eIF-2）和真核起始因子2B（eIF-2B）表达的调节表明，GCN4的翻译与eIF-2·GTP·甲硫氨酰-tRNA<sup>(iMet)</sup>三元复合物的水平呈负相关。

Modulation of tRNA(iMet), eIF-2, and eIF-2B expression shows that GCN4 translation is inversely coupled to the level of eIF-2.GTP.Met-tRNA(iMet) ternary complexes.

作者信息

Dever T E, Yang W, Aström S, Byström A S, Hinnebusch A G

机构信息

Section on Molecular Genetics of Lower Eukaryotes, National Institute of Child Health and Human Development, Bethesda, Maryland 20892-2785, USA.

出版信息

Mol Cell Biol. 1995 Nov;15(11):6351-63. doi: 10.1128/MCB.15.11.6351.

DOI:10.1128/MCB.15.11.6351
PMID:7565788
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC230887/
Abstract

To understand how phosphorylation of eukaryotic translation initiation factor (eIF)-2 alpha in Saccharomyces cerevisiae stimulates GCN4 mRNA translation while at the same time inhibiting general translation initiation, we examined the effects of altering the gene dosage of initiator tRNA(Met), eIF-2, and the guanine nucleotide exchange factor for eIF-2, eIF-2B. Overexpression of all three subunits of eIF-2 or all five subunits of eIF-2B suppressed the effects of eIF-2 alpha hyperphosphorylation on both GCN4-specific and general translation initiation. Consistent with eIF-2 functioning in translation as part of a ternary complex composed of eIF-2, GTP, and Met-tRNA(iMet), reduced gene dosage of initiator tRNA(Met) mimicked phosphorylation of eIF-2 alpha and stimulated GCN4 translation. In addition, overexpression of a combination of eIF-2 and tRNA(iMet) suppressed the growth-inhibitory effects of eIF-2 hyperphosphorylation more effectively than an increase in the level of either component of the ternary complex alone. These results provide in vivo evidence that phosphorylation of eIF-2 alpha reduces the activities of both eIF-2 and eIF-2B and that the eIF-2.GTP. Met-tRNA(iMet) ternary complex is the principal component limiting translation in cells when eIF-2 alpha is phosphorylated on serine 51. Analysis of eIF-2 alpha phosphorylation in the eIF-2-overexpressing strain also provides in vivo evidence that phosphorylated eIF-2 acts as a competitive inhibitor of eIF-2B rather than forming an excessively stable inactive complex. Finally, our results demonstrate that the concentration of eIF-2-GTP. Met-tRNA(iMet) ternary complexes is the cardinal parameter determining the site of reinitiation on GCN4 mRNA and support the idea that reinitiation at GCN4 is inversely related to the concentration of ternary complexes in the cell.

摘要

为了解酿酒酵母中真核生物翻译起始因子（eIF）-2α的磷酸化如何刺激GCN4 mRNA的翻译，同时抑制一般的翻译起始，我们研究了改变起始tRNA（Met）、eIF-2以及eIF-2的鸟嘌呤核苷酸交换因子eIF-2B的基因剂量所产生的影响。eIF-2的所有三个亚基或eIF-2B的所有五个亚基的过表达均抑制了eIF-2α过度磷酸化对GCN4特异性翻译起始和一般翻译起始的影响。与eIF-2作为由eIF-2、GTP和Met-tRNA（iMet）组成的三元复合物的一部分在翻译中发挥作用一致，起始tRNA（Met）基因剂量的减少模拟了eIF-2α的磷酸化并刺激了GCN4的翻译。此外，eIF-2和tRNA（iMet）组合的过表达比单独增加三元复合物中任何一种成分的水平更有效地抑制了eIF-2过度磷酸化对生长的抑制作用。这些结果提供了体内证据，表明eIF-2α的磷酸化降低了eIF-2和eIF-2B的活性，并且当eIF-2α在丝氨酸51处被磷酸化时，eIF-2·GTP·Met-tRNA（iMet）三元复合物是限制细胞内翻译的主要成分。对eIF-2过表达菌株中eIF-2α磷酸化的分析还提供了体内证据，表明磷酸化的eIF-2作为eIF-2B的竞争性抑制剂，而不是形成过度稳定的无活性复合物。最后，我们的结果表明eIF-2-GTP·Met-tRNA（iMet）三元复合物的浓度是决定GCN4 mRNA重新起始位点的关键参数，并支持这样一种观点，即GCN4处的重新起始与细胞中三元复合物的浓度呈负相关。