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PMID: 2390971 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Cloning and expression of eukaryotic initiation factor 4B cDNA: sequence determination identifies a common RNA recognition motif.

The EMBO journal ·Vol. 9 ·No. 9 ·1990-09-00 ·Pages 2783-90

Milburn SC, Hershey JW, Davies MV, Kelleher K, Kaufman RJ

Abstract

Eukaryotic protein synthesis initiation factor 4B (eIF-4B) is an 80,000 dalton polypeptide which is essential for the binding of mRNA to ribosomes. A highly purified preparation of eIF-4B from HeLa cells was subjected to enzymatic cleavage and amino-terminal amino acid sequence analysis. Degenerate oligonucleotide probes were used to isolate a 3851 bp cDNA encoding eIF-4B from a human cDNA library. The DNA encodes a protein comprising 611 residues with a mass of 69,843 daltons. The amino-terminal domain of eIF-4B contains a consensus RNA binding domain present in a number of other RNA binding proteins. Expression of eIF-4B in transfected COS-1 cells yielded a polypeptide which reacted with anti-eIF-4B antiserum and comigrated with purified eIF-4B. Expression of eIF-4B in COS-1 cells resulted in a general inhibition of translation, possibly due to a 50-fold eIF-4B overproduction.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Binding Sites Cell Line Cloning, Molecular Eukaryotic Initiation Factors Gene Expression HeLa Cells/metabolism Humans Molecular Sequence Data Oligonucleotide Probes Peptide Fragments/isolation & purification Peptide Initiation Factors/genetics Protein Conformation RNA, Messenger/genetics,metabolism Sequence Homology, Nucleic Acid Transfection
Chemicals
Eukaryotic Initiation Factors Oligonucleotide Probes Peptide Fragments Peptide Initiation Factors RNA, Messenger eIF-4B
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Milburn S C
Genetics Institute, Cambridge, MA 02140.
Hershey J W
Davies M V
Kelleher K
Kaufman R J
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1990-09-00
Pages
2783-90
Language
English
Region
England
NLM ID
8208664
PMCID
PMC551988
Subset
IM
Grants
NIGMS NIH HHS · GM22135 · United States
External Links
PubMed source PMC Full Text View journal details
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