Document Detail


Cloning and expression of cDNAs for the beta subunit of eukaryotic initiation factor-2B, the guanine nucleotide exchange factor for eukaryotic initiation factor-2.
MedLine Citation:
PMID:  7639677     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A key control point in the initiation of protein synthesis in mammalian cells is the recycling of eukaryotic initiation factor (eIF)-2 by the guanine nucleotide exchange factor eIF-2B. In mammalian cells, eIF-2B is a complex of five different subunits termed epsilon, delta, gamma, beta and alpha. To clone cDNAs for the beta subunit of rabbit eIF-2B, amino acid sequence data was first obtained and used to design redundant oligonucleotide primers for use in PCR. PCR products were used to screen a rabbit liver cDNA library in lambda gt11 to obtain full-length cDNAs for eIF-2B beta. The cDNAs were sequenced completely on both strands and revealed an open reading frame encoding a predicted 351-amino acid polypeptide of 39.0 kDa. The molecular mass and pI (5.99) of the predicted protein agree well with the properties of eIF-2B beta purified from rabbit reticulocytes. In vitro transcription/-translation of the cDNAs gave rise to a product that migrated at a position indistinguishable from that of this subunit of the purified protein. The amino acid sequence shows a high degree of similarity to that of GCD7, a Saccharomyces cerevisiae protein thought to be equivalent to mammalian eIF-2B beta. Northern-blot analysis revealed a single major mRNA species for eIF-2B beta in each of the four rabbit tissues tested.
Authors:
B L Craddock; N T Price; C G Proud
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Biochemical journal     Volume:  309 ( Pt 3)     ISSN:  0264-6021     ISO Abbreviation:  Biochem. J.     Publication Date:  1995 Aug 
Date Detail:
Created Date:  1995-09-13     Completed Date:  1995-09-13     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  1009-14     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, School of Medical Sciences, University of Bristol U.K.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/Z48222
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Base Sequence
Blotting, Northern
Cloning, Molecular
DNA, Complementary
Eukaryotic Initiation Factor-2 / metabolism*
Guanine Nucleotide Exchange Factors
Molecular Sequence Data
Proteins / genetics*,  metabolism
RNA, Messenger / genetics,  metabolism
Rabbits
Sequence Homology, Amino Acid
Grant Support
ID/Acronym/Agency:
//Wellcome Trust
Chemical
Reg. No./Substance:
0/DNA, Complementary; 0/Eukaryotic Initiation Factor-2; 0/Guanine Nucleotide Exchange Factors; 0/Proteins; 0/RNA, Messenger
Comments/Corrections

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