Home LiteratureArticle Details
PMID: 8417348 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

GCD11, a negative regulator of GCN4 expression, encodes the gamma subunit of eIF-2 in Saccharomyces cerevisiae.

Molecular and cellular biology ·Vol. 13 ·No. 1 ·1993-01-00 ·Pages 506-20

Hannig EM, Cigan AM, Freeman BA, Kinzy TG

Abstract

The eukaryotic translation initiation factor eIF-2 plays a critical role in regulating the expression of the yeast transcriptional activator GCN4. Mutations in genes encoding the alpha and beta subunits of eIF-2 alter translational efficiency at the GCN4 AUG codon and constitutively elevate GCN4 translation. Mutations in the yeast GCD11 gene have been shown to confer a similar phenotype. The nucleotide sequence of the cloned GCD11 gene predicts a 527-amino-acid polypeptide that is similar to the prokaryotic translation elongation factor EF-Tu. Relative to EF-Tu, the deduced GCD11 amino acid sequence contains a 90-amino-acid N-terminal extension and an internal cysteine-rich sequence that contains a potential metal-binding finger motif. We have identified the GCD11 gene product as the gamma subunit of eIF-2 by the following criteria: (i) sequence identities with mammalian eIF-2 gamma peptides; (ii) increased eIF-2 activity in extracts prepared from cells cooverexpressing GCD11, eIF-2 alpha, and eIF-2 beta; and (iii) cross-reactivity of antibodies directed against the GCD11 protein with the 58-kDa polypeptide present in purified yeast eIF-2. The predicted GCD11 polypeptide contains all of the consensus elements known to be required for guanine nucleotide binding, suggesting that, in Saccharomyces cerevisiae, the gamma subunit of eIF-2 is responsible for GDP-GTP binding.

Related Genes
EF-Tu GCD11 GCN4 GST1
MeSH Terms
Amino Acid Sequence Animals Base Sequence Cloning, Molecular Cross Reactions DNA, Fungal/genetics DNA-Binding Proteins Eukaryotic Initiation Factor-2/chemistry,genetics,immunology Fungal Proteins/genetics GTP-Binding Proteins/genetics,metabolism Gene Expression Gene Expression Regulation, Fungal Genes, Fungal Macromolecular Substances Molecular Sequence Data Oligodeoxyribonucleotides/chemistry Peptide Elongation Factor Tu/genetics Protein Kinases/genetics RNA, Fungal/genetics RNA, Messenger/genetics Rabbits Restriction Mapping Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Sequence Alignment Swine Transcription Factors/genetics
Chemicals
DNA, Fungal DNA-Binding Proteins Eukaryotic Initiation Factor-2 Fungal Proteins Macromolecular Substances Oligodeoxyribonucleotides RNA, Fungal RNA, Messenger Saccharomyces cerevisiae Proteins Transcription Factors Protein Kinases GTP-Binding Proteins Peptide Elongation Factor Tu
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Hannig E M
Molecular and Cell Biology Program, University of Texas, Dallas 830688.
Cigan A M
Freeman B A
Kinzy T G
References (67)
67 references, click to expand
  1. A hierarchy of trans-acting factors modulates translation of an activator of amino acid biosynthetic genes in Saccharomyces cerevisiae.
    Mol Cell Biol. 1985 Sep;5(9):2349-60 PMID: 3915540
  2. Sigma factors from E. coli, B. subtilis, phage SP01, and phage T4 are homologous proteins.
    Nucleic Acids Res. 1986 Aug 26;14(16):6745-63 PMID: 3092189
  3. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector.
    Gene. 1987;60(2-3):237-43 PMID: 3327750
  4. Multiple GCD genes required for repression of GCN4, a transcriptional activator of amino acid biosynthetic genes in Saccharomyces cerevisiae.
    Mol Cell Biol. 1986 Nov;6(11):3990-8 PMID: 3540603
  5. Structure of the two genes coding for polypeptide chain elongation factor 1 alpha (EF-1 alpha) from Saccharomyces cerevisiae.
    Gene. 1986;45(3):265-73 PMID: 3026912
  6. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes.
    J Biol Chem. 1987 Jul 25;262(21):10035-8 PMID: 3611052
  7. Interactions between positive and negative regulators of GCN4 controlling gene expression and entry into the yeast cell cycle.
    Genetics. 1987 Nov;117(3):409-19 PMID: 3319768
  8. A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli.
    Gene. 1987;57(2-3):267-72 PMID: 3319781
  9. A yeast gene required for the G1-to-S transition encodes a protein containing an A-kinase target site and GTPase domain.
    EMBO J. 1988 Apr;7(4):1175-82 PMID: 2841115
  10. Mutations at a Zn(II) finger motif in the yeast eIF-2 beta gene alter ribosomal start-site selection during the scanning process.
    Cell. 1988 Aug 26;54(5):621-32 PMID: 3136928
  11. Structure of the beta subunit of translational initiation factor eIF-2.
    Cell. 1988 Aug 26;54(5):633-9 PMID: 3044606
  12. Mechanisms of gene regulation in the general control of amino acid biosynthesis in Saccharomyces cerevisiae.
    Microbiol Rev. 1988 Jun;52(2):248-73 PMID: 3045517
  13. tRNAi(met) functions in directing the scanning ribosome to the start site of translation.
    Science. 1988 Oct 7;242(4875):93-7 PMID: 3051379
  14. Molecular analysis of GCN3, a translational activator of GCN4: evidence for posttranslational control of GCN3 regulatory function.
    Mol Cell Biol. 1988 Nov;8(11):4808-20 PMID: 3062370
  15. The scanning model for translation: an update.
    J Cell Biol. 1989 Feb;108(2):229-41 PMID: 2645293
  16. Yeast translation initiation suppressor sui2 encodes the alpha subunit of eukaryotic initiation factor 2 and shares sequence identity with the human alpha subunit.
    Proc Natl Acad Sci U S A. 1989 Apr;86(8):2784-8 PMID: 2649894
  17. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  18. Structure of the guanine-nucleotide-binding domain of the Ha-ras oncogene product p21 in the triphosphate conformation.
    Nature. 1989 Sep 21;341(6239):209-14 PMID: 2476675
  19. Sequences that surround the stop codons of upstream open reading frames in GCN4 mRNA determine their distinct functions in translational control.
    Genes Dev. 1989 Aug;3(8):1217-25 PMID: 2676723
  20. Mutations in the structural genes for eukaryotic initiation factors 2 alpha and 2 beta of Saccharomyces cerevisiae disrupt translational control of GCN4 mRNA.
    Proc Natl Acad Sci U S A. 1989 Oct;86(19):7515-9 PMID: 2678106
  21. Identification of positive-acting domains in GCN2 protein kinase required for translational activation of GCN4 expression.
    Mol Cell Biol. 1990 Jun;10(6):2820-31 PMID: 2188100
  22. Characteristics of eukaryotic initiation factor 2 and its subunits.
    J Biol Chem. 1980 Feb 10;255(3):1189-93 PMID: 6153180
  23. Affinity labeling of eukaryotic initiation factor 2 and elongation factor 1 alpha beta gamma with GTP analogs.
    Arch Biochem Biophys. 1990 Aug 15;281(1):157-62 PMID: 2383020
  24. Initiation of mRNA translation in prokaryotes.
    Biochemistry. 1990 Jun 26;29(25):5881-9 PMID: 2200518
  25. The translational activator GCN3 functions downstream from GCN1 and GCN2 in the regulatory pathway that couples GCN4 expression to amino acid availability in Saccharomyces cerevisiae.
    Genetics. 1990 Nov;126(3):549-62 PMID: 2249755
  26. Suppression of ribosomal reinitiation at upstream open reading frames in amino acid-starved cells forms the basis for GCN4 translational control.
    Mol Cell Biol. 1991 Jan;11(1):486-96 PMID: 1986242
  27. The GTPase superfamily: conserved structure and molecular mechanism.
    Nature. 1991 Jan 10;349(6305):117-27 PMID: 1898771
  28. Chemical modification of pig liver initiation factor eIF-2 with N-ethylmaleimide. Amino acid sequences around the N-ethylmaleimide-reactive sulfhydryl groups and the effect of GDP on the modification.
    J Biochem. 1990 Oct;108(4):635-41 PMID: 2292592
  29. Divergence and conservation of SUP2 (SUP35) gene of yeast Pichia pinus and Saccharomyces cerevisiae.
    Yeast. 1990 Nov-Dec;6(6):461-72 PMID: 2080663
  30. Complex formation by positive and negative translational regulators of GCN4.
    Mol Cell Biol. 1991 Jun;11(6):3217-28 PMID: 2038327
  31. Translational control in mammalian cells.
    Annu Rev Biochem. 1991;60:717-55 PMID: 1883206
  32. Amino acid sequence analysis of the beta- and gamma-subunits of eukaryotic initiation factor eIF-2. Identification of regions interacting with GTP.
    Biochim Biophys Acta. 1991 Sep 20;1079(3):308-15 PMID: 1911855
  33. Saccharomyces cerevisiae elongation factor 2. Genetic cloning, characterization of expression, and G-domain modeling.
    J Biol Chem. 1992 Jan 15;267(2):1190-7 PMID: 1730643
  34. A model for the aminoacyl-tRNA binding site of eukaryotic elongation factor 1 alpha.
    J Biol Chem. 1992 Jan 25;267(3):1623-32 PMID: 1730707
  35. The structure of Ras protein: a model for a universal molecular switch.
    Trends Biochem Sci. 1991 Oct;16(10):382-7 PMID: 1785141
  36. Phosphorylation of initiation factor 2 alpha by protein kinase GCN2 mediates gene-specific translational control of GCN4 in yeast.
    Cell. 1992 Feb 7;68(3):585-96 PMID: 1739968
  37. Refined structure of elongation factor EF-Tu from Escherichia coli.
    J Mol Biol. 1992 Feb 5;223(3):721-42 PMID: 1542116
  38. Mechanism and regulation of eukaryotic protein synthesis.
    Microbiol Rev. 1992 Jun;56(2):291-315 PMID: 1620067
  39. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  40. Detection of specific sequences among DNA fragments separated by gel electrophoresis.
    J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
  41. High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules.
    Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035-9 PMID: 375221
  42. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments.
    Gene. 1979 Dec;8(1):17-24 PMID: 395030
  43. DNA sequences from the str operon of Escherichia coli.
    J Biol Chem. 1980 May 25;255(10):4660-6 PMID: 6989816
  44. The complete amino-acid sequence of elongation factor Tu from Escherichia coli.
    Eur J Biochem. 1980 Jul;108(2):507-26 PMID: 6997043
  45. The nucleotide sequence of the cloned tufA gene of Escherichia coli.
    Gene. 1980 Dec;12(1-2):25-31 PMID: 7011903
  46. The nucleotide sequence of tufB and four nearby tRNA structural genes of Escherichia coli.
    Gene. 1980 Dec;12(1-2):33-9 PMID: 7011904
  47. Physical analysis of mating-type loci in Saccharomyces cerevisiae.
    Cold Spring Harb Symp Quant Biol. 1981;45 Pt 2:961-81 PMID: 7021055
  48. The primary structure of elongation factor G from Escherichia coli. A complete amino acid sequence.
    FEBS Lett. 1982 Mar 8;139(1):130-5 PMID: 7042386
  49. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  50. Repeated DNA sequences upstream from HIS1 also occur at several other co-regulated genes in Saccharomyces cerevisiae.
    J Biol Chem. 1983 Apr 25;258(8):5238-47 PMID: 6300123
  51. New initiation factor activity required for globin mRNA translation.
    J Biol Chem. 1983 May 10;258(9):5804-10 PMID: 6853548
  52. The histidine tRNA genes of yeast.
    J Biol Chem. 1983 Jul 10;258(13):8175-82 PMID: 6305975
  53. One-step gene disruption in yeast.
    Methods Enzymol. 1983;101:202-11 PMID: 6310324
  54. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
    Anal Biochem. 1983 Jul 1;132(1):6-13 PMID: 6312838
  55. A comprehensive set of sequence analysis programs for the VAX.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387-95 PMID: 6546423
  56. A comprehensive sequence analysis program for the IBM personal computer.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):581-99 PMID: 6546431
  57. Saccharomyces cerevisiae killer virus transcripts contain template-coded polyadenylate tracts.
    Mol Cell Biol. 1984 Jan;4(1):101-9 PMID: 6199660
  58. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing.
    Gene. 1984 Jun;28(3):351-9 PMID: 6235151
  59. Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Aug;4(8):1440-8 PMID: 6092912
  60. Positive regulatory interactions of the HIS4 gene of Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Jul;4(7):1326-33 PMID: 6095062
  61. Sequence of the initiation factor IF2 gene: unusual protein features and homologies with elongation factors.
    Proc Natl Acad Sci U S A. 1984 Dec;81(24):7787-91 PMID: 6096856
  62. Assembly of the mitochondrial membrane system. CBP6, a yeast nuclear gene necessary for synthesis of cytochrome b.
    J Biol Chem. 1985 Feb 10;260(3):1513-20 PMID: 2981859
  63. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance.
    Mol Gen Genet. 1984;197(2):345-6 PMID: 6394957
  64. Identification of two genes coding for the translation elongation factor EF-1 alpha of S. cerevisiae.
    EMBO J. 1984 Dec 20;3(13):3311-5 PMID: 6396088
  65. Cloning, nucleotide sequence, and expression of one of two genes coding for yeast elongation factor 1 alpha.
    J Biol Chem. 1985 Mar 10;260(5):3090-6 PMID: 2982849
  66. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  67. Potential metal-binding domains in nucleic acid binding proteins.
    Science. 1986 Apr 25;232(4749):485-7 PMID: 2421409
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.13.1.506-520.1993
Published
1993-01-00
Pages
506-20
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358930
Subset
IM
Grants
NIGMS NIH HHS · GM26796 · United States
Databases
GENBANK
L04268
External Links
PubMed source DOI: 10.1128/mcb.13.1.506-520.1993 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2