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PMID: 8441423 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Evidence that GCD6 and GCD7, translational regulators of GCN4, are subunits of the guanine nucleotide exchange factor for eIF-2 in Saccharomyces cerevisiae.

Molecular and cellular biology ·Vol. 13 ·No. 3 ·1993-03-00 ·Pages 1920-32

Bushman JL, Asuru AI, Matts RL, Hinnebusch AG

Abstract

Starvation of the yeast Saccharomyces cerevisiae for an amino acid signals increased translation of GCN4, a transcriptional activator of amino acid biosynthetic genes. We have isolated and characterized the GCD6 and GCD7 genes and shown that their products are required to repress GCN4 translation under nonstarvation conditions. We find that both GCD6 and GCD7 show sequence similarities to components of a high-molecular-weight complex (the GCD complex) that appears to be the yeast equivalent of translation initiation factor 2B (eIF-2B), which catalyzes GDP-GTP exchange on eIF-2. Furthermore, we show that GCD6 is 30% identical to the largest subunit of eIF-2B isolated from rabbit reticulocytes. Deletion of either GCD6 or GCD7 is lethal, and nonlethal mutations in these genes increase GCN4 translation in the same fashion described for defects in known subunits of eIF-2 or the GCD complex; derepression of GCN4 is dependent on short open reading frames in the GCN4 mRNA leader and occurs independently of eIF-2 alpha phosphorylation by protein kinase GCN2, which is normally required to stimulate GCN4 translation. Together, our results provide evidence that GCD6 and GCD7 are subunits of eIF-2B in S. cerevisiae and further implicate this GDP-GTP exchange factor in gene-specific translational control.

MeSH Terms
Amino Acid Sequence Amino Acids/biosynthesis Animals Base Sequence DNA-Binding Proteins Eukaryotic Initiation Factor-2/metabolism Eukaryotic Initiation Factor-2B Fungal Proteins/biosynthesis,genetics,metabolism Gene Expression Regulation, Fungal Guanine Nucleotide Exchange Factors Macromolecular Substances Molecular Sequence Data Mutation Peptide Initiation Factors/biosynthesis Protein Biosynthesis Protein Conformation Protein Kinases/biosynthesis,metabolism Proteins/chemistry,genetics Rabbits Repressor Proteins/genetics Reticulocytes/chemistry Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid
Chemicals
Amino Acids DNA-Binding Proteins Eukaryotic Initiation Factor-2 Eukaryotic Initiation Factor-2B Fungal Proteins GCD6 protein, S cerevisiae GCD7 protein, S cerevisiae Guanine Nucleotide Exchange Factors Macromolecular Substances Peptide Initiation Factors Proteins Repressor Proteins Saccharomyces cerevisiae Proteins Protein Kinases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Bushman J L
Section on Molecular Genetics of Lower Eukaryotes, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.
Asuru A I
Matts R L
Hinnebusch A G
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
Published
1993-03-00
Pages
1920-32
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359506
Subset
IM
Grants
NIEHS NIH HHS · ES-04299 · United States
Databases
GENBANK
L07115, L07116
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