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Cloning and expression of eukaryotic initiation factor 4B cDNA: sequence determination identifies a common RNA recognition motif.
Author(s) -
Milburn S. C.,
Hershey J. W.,
Davies M. V.,
Kelleher K.,
Kaufman R. J.
Publication year - 1990
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1990.tb07466.x
Subject(s) - biology , cloning (programming) , complementary dna , genetics , sequence motif , computational biology , motif (music) , rna , gene , computer science , physics , acoustics , programming language
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.