| Cloning and expression of cDNAs for the beta subunit of eukaryotic initiation factor-2B, the guanine nucleotide exchange factor for eukaryotic initiation factor-2. | |
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MedLine Citation:
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PMID: 7639677 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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B L Craddock; N T Price; C G Proud |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: The Biochemical journal Volume: 309 ( Pt 3) ISSN: 0264-6021 ISO Abbreviation: Biochem. J. Publication Date: 1995 Aug |
Date Detail:
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Created Date: 1995-09-13 Completed Date: 1995-09-13 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 2984726R Medline TA: Biochem J Country: ENGLAND |
Other Details:
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Languages: eng Pagination: 1009-14 Citation Subset: IM |
Affiliation:
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Department of Biochemistry, School of Medical Sciences, University of Bristol U.K. |
| Data Bank Information | |
Bank Name/Acc. No.:
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GENBANK/Z48222 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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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:
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//Wellcome Trust |
| Chemical | |
Reg. No./Substance:
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0/DNA, Complementary; 0/Eukaryotic Initiation Factor-2; 0/Guanine Nucleotide Exchange Factors; 0/Proteins; 0/RNA, Messenger |
| Comments/Corrections | |
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