Budding Yeast GCN1 Binds the GI Domain to Activate the eIF2α Kinase GCN2

Hiroyuki Kubota, Kazuhisa Ota, Yoshiyuki Sakaki, Takashi Ito

研究成果: ジャーナルへの寄稿記事

37 引用 (Scopus)

抄録

When starved for a single amino acid, the budding yeast Saccharomyces cerevisiae activates the eukaryotic initiation factor 2α (eIF2α) kinase GCN2 in a GCN1-dependent manner. Phosphorylated eIF2α inhibits general translation but selectively derepresses the synthesis of the transcription factor GCN4, which leads to coordinated induction of genes involved in biosynthesis of various amino acids, a phenomenon called general control response. We recently demonstrated that this response requires binding of GCN1 to the GI domain occurring at the N terminus of GCN2 (Kubota, H., Sakaki, Y., and Ito, T. (2000) J. Biol. Chem. 275, 20243-20246). Here we provide the first evidence for the involvement of GCN1-GCN2 interaction in activation of GCN2 per se. We identified a C-terminal segment of GCN1 sufficient to bind the GI domain and used a novel dual bait two-hybrid method to identify mutations rendering GCN1 incapable of interacting with GCN2. The yeast bearing such an allele, gcn1-F2291L, fails to display derepression of GCN4 translation and hence general control response, as does a GI domain mutant, gcn2-Y74A, defective in association with GCN1. Furthermore, we demonstrated that phosphorylation of eIF2α is impaired in both mutants. Since GCN2 is the sole eIF2α kinase in yeast, these findings indicate a critical role of GCN1-GCN2 interaction in activation of the kinase in vivo.

元の言語英語
ページ(範囲)17591-17596
ページ数6
ジャーナルJournal of Biological Chemistry
276
発行部数20
DOI
出版物ステータス出版済み - 5 18 2001
外部発表Yes

Fingerprint

Eukaryotic Initiation Factor-2
Saccharomycetales
Yeast
Phosphotransferases
Bearings (structural)
Yeasts
Chemical activation
Amino Acids
Two-Hybrid System Techniques
Phosphorylation
Biosynthesis
Saccharomyces cerevisiae
Transcription Factors
Genes
Alleles
Association reactions
Mutation

All Science Journal Classification (ASJC) codes

  • Biochemistry

これを引用

Budding Yeast GCN1 Binds the GI Domain to Activate the eIF2α Kinase GCN2. / Kubota, Hiroyuki; Ota, Kazuhisa; Sakaki, Yoshiyuki; Ito, Takashi.

:: Journal of Biological Chemistry, 巻 276, 番号 20, 18.05.2001, p. 17591-17596.

研究成果: ジャーナルへの寄稿記事

Kubota, Hiroyuki ; Ota, Kazuhisa ; Sakaki, Yoshiyuki ; Ito, Takashi. / Budding Yeast GCN1 Binds the GI Domain to Activate the eIF2α Kinase GCN2. :: Journal of Biological Chemistry. 2001 ; 巻 276, 番号 20. pp. 17591-17596.
@article{88f6c44d6bed497fb8537956915f0f3c,
title = "Budding Yeast GCN1 Binds the GI Domain to Activate the eIF2α Kinase GCN2",
abstract = "When starved for a single amino acid, the budding yeast Saccharomyces cerevisiae activates the eukaryotic initiation factor 2α (eIF2α) kinase GCN2 in a GCN1-dependent manner. Phosphorylated eIF2α inhibits general translation but selectively derepresses the synthesis of the transcription factor GCN4, which leads to coordinated induction of genes involved in biosynthesis of various amino acids, a phenomenon called general control response. We recently demonstrated that this response requires binding of GCN1 to the GI domain occurring at the N terminus of GCN2 (Kubota, H., Sakaki, Y., and Ito, T. (2000) J. Biol. Chem. 275, 20243-20246). Here we provide the first evidence for the involvement of GCN1-GCN2 interaction in activation of GCN2 per se. We identified a C-terminal segment of GCN1 sufficient to bind the GI domain and used a novel dual bait two-hybrid method to identify mutations rendering GCN1 incapable of interacting with GCN2. The yeast bearing such an allele, gcn1-F2291L, fails to display derepression of GCN4 translation and hence general control response, as does a GI domain mutant, gcn2-Y74A, defective in association with GCN1. Furthermore, we demonstrated that phosphorylation of eIF2α is impaired in both mutants. Since GCN2 is the sole eIF2α kinase in yeast, these findings indicate a critical role of GCN1-GCN2 interaction in activation of the kinase in vivo.",
author = "Hiroyuki Kubota and Kazuhisa Ota and Yoshiyuki Sakaki and Takashi Ito",
year = "2001",
month = "5",
day = "18",
doi = "10.1074/jbc.M011793200",
language = "English",
volume = "276",
pages = "17591--17596",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "20",

}

TY - JOUR

T1 - Budding Yeast GCN1 Binds the GI Domain to Activate the eIF2α Kinase GCN2

AU - Kubota, Hiroyuki

AU - Ota, Kazuhisa

AU - Sakaki, Yoshiyuki

AU - Ito, Takashi

PY - 2001/5/18

Y1 - 2001/5/18

N2 - When starved for a single amino acid, the budding yeast Saccharomyces cerevisiae activates the eukaryotic initiation factor 2α (eIF2α) kinase GCN2 in a GCN1-dependent manner. Phosphorylated eIF2α inhibits general translation but selectively derepresses the synthesis of the transcription factor GCN4, which leads to coordinated induction of genes involved in biosynthesis of various amino acids, a phenomenon called general control response. We recently demonstrated that this response requires binding of GCN1 to the GI domain occurring at the N terminus of GCN2 (Kubota, H., Sakaki, Y., and Ito, T. (2000) J. Biol. Chem. 275, 20243-20246). Here we provide the first evidence for the involvement of GCN1-GCN2 interaction in activation of GCN2 per se. We identified a C-terminal segment of GCN1 sufficient to bind the GI domain and used a novel dual bait two-hybrid method to identify mutations rendering GCN1 incapable of interacting with GCN2. The yeast bearing such an allele, gcn1-F2291L, fails to display derepression of GCN4 translation and hence general control response, as does a GI domain mutant, gcn2-Y74A, defective in association with GCN1. Furthermore, we demonstrated that phosphorylation of eIF2α is impaired in both mutants. Since GCN2 is the sole eIF2α kinase in yeast, these findings indicate a critical role of GCN1-GCN2 interaction in activation of the kinase in vivo.

AB - When starved for a single amino acid, the budding yeast Saccharomyces cerevisiae activates the eukaryotic initiation factor 2α (eIF2α) kinase GCN2 in a GCN1-dependent manner. Phosphorylated eIF2α inhibits general translation but selectively derepresses the synthesis of the transcription factor GCN4, which leads to coordinated induction of genes involved in biosynthesis of various amino acids, a phenomenon called general control response. We recently demonstrated that this response requires binding of GCN1 to the GI domain occurring at the N terminus of GCN2 (Kubota, H., Sakaki, Y., and Ito, T. (2000) J. Biol. Chem. 275, 20243-20246). Here we provide the first evidence for the involvement of GCN1-GCN2 interaction in activation of GCN2 per se. We identified a C-terminal segment of GCN1 sufficient to bind the GI domain and used a novel dual bait two-hybrid method to identify mutations rendering GCN1 incapable of interacting with GCN2. The yeast bearing such an allele, gcn1-F2291L, fails to display derepression of GCN4 translation and hence general control response, as does a GI domain mutant, gcn2-Y74A, defective in association with GCN1. Furthermore, we demonstrated that phosphorylation of eIF2α is impaired in both mutants. Since GCN2 is the sole eIF2α kinase in yeast, these findings indicate a critical role of GCN1-GCN2 interaction in activation of the kinase in vivo.

UR - http://www.scopus.com/inward/record.url?scp=0035907288&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035907288&partnerID=8YFLogxK

U2 - 10.1074/jbc.M011793200

DO - 10.1074/jbc.M011793200

M3 - Article

C2 - 11350982

AN - SCOPUS:0035907288

VL - 276

SP - 17591

EP - 17596

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 20

ER -