NDLI: Molecular cloning and characterization of cDNA encoding the alpha subunit of the rat protein synthesis initiation factor eIF-2B.

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Molecular cloning and characterization of cDNA encoding the alpha subunit of the rat protein synthesis initiation factor eIF-2B.

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Molecular cloning and characterization of cDNA encoding the alpha subunit of the rat protein synthesis initiation factor eIF-2B.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Feldhoff, R. C. Hinnebusch, A. G. Kimball, S. R. Jefferson, L. S. Flowers, K. M.
Description	Author Affiliation: Flowers KM ( Department of Cellular and Molecular Physiology, College of Medicine, Pennsylvania State University, Hershey 17033, USA.);
Abstract	Eukaryotic initiation factor 2B (eIF-2B) is an essential component of the pathway of peptide-chain initiation in mammalian cells, yet little is known about its molecular structure and regulation. To investigate the structure, regulation, and interactions of the individual subunits of eIF-2B, we have begun to clone, characterize, and express the corresponding cDNAs. We report here the cloning and characterization of a 1510-bp cDNA encoding the alpha subunit of eIF-2B from a rat brain cDNA library. The cDNA contains an open reading frame of 918 bp encoding a polypeptide of 305 aa with a predicted molecular mass of 33.7 kDa. This cDNA recognizes a single RNA species approximately 1.6 kb in length on Northern blots of RNA from rat liver. The predicted amino acid sequence contains regions identical to the sequences of peptides derived from bovine liver eIF-2B alpha subunit. Expression of this cDNA in vitro yields a peptide which comigrates with natural eIF-2B alpha in SDS/polyacrylamide gels. The predicted amino acid sequence exhibits 42% identity to that deduced for the Saccharomyces cerevisiae GCN3 protein, the smallest subunit of yeast eIF-2B. In addition, expression of the rat cDNA in yeast functionally complements a gcn3 deletion for the inability to induce histidine biosynthetic genes under the control of GCN4. These results strongly support the hypothesis that mammalian eIF-2 alpha and GCN3 are homologues. Southern blots indicate that the eIF-2B alpha cDNA also recognizes genomic DNA fragments from several other species, suggesting significant homology between the rat eIF-2B alpha gene and that from other species.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	10
Volume Number	92
Language	English
Publisher	National Academy of Sciences
Publisher Date	1995-06-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Brain Metabolism Protein Biosynthesis Amino Acid Sequence Animals Blotting, Southern Cloning, Molecular DNA Primers DNA, Complementary Drosophila Melanogaster Gene Expression Gene Library Genetic Complementation Test Guanine Nucleotide Exchange Factors Macromolecular Substances Mice Molecular Sequence Data Polymerase Chain Reaction Proteins Chemistry Isolation & Purification Rabbits Saccharomyces Cerevisiae Sequence Homology, Amino Acid Comparative Study Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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