Purification and characterization of a new eukaryotic protein translation factor. Eukaryotic initiation factor 4H.
A new protein with translational activity has been identified on the basis of its ability to stimulate translation in an in vitro globin synthesis assay deficient in eukaryotic initiation factor (eIF) 4B and eIF4F. This protein has been purified to greater than 80% homogeneity from rabbit reticulocyte lysate and has been given the name eIF4H. eIF4H was shown to stimulate the in vitro activities of eIF4B and eIF4F in globin synthesis, as well as the in vitro RNA-dependent ATPase activities of eIF4A, eIF4B, and eIF4F. Three tryptic fragments of eIF4H yielded amino acid sequences that were 100% identical to a human sequence found in the GeneBankTM that codes for a previously uncharacterized protein (HUMORFU_1). The calculated molecular weight of the protein encoded by this sequence, its predicted cyanogen bromide fragmentation, and calculated isoelectric point are all consistent with those determined experimentally for eIF4H. Also, the presence of an RNA recognition motif within HUMORFU_1 suggests that eIF4H may interact with mRNA. We conclude that this newly characterized protein, eIF4H, functions to stimulate the initiation of protein synthesis at the level of mRNA utilization, and is encoded by the gene for HUMORFU_1.
Richter-Cook
N J
NJ
Departments of Biochemistry, Case Western Reserve University, Cleveland, Ohio 44106-4935, USA.
Dever
T E
TE
Hensold
J O
JO
Merrick
W C
WC
D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D039642:Eukaryotic Initiation Factors; D006801:Humans; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010448:Peptide Initiation Factors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D010449:Peptide Mapping; D014176:Protein Biosynthesis; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D011817:Rabbits; D012156:Reticulocytes / Q000737:chemistry; D016415:Sequence Alignment
C111495:EIF4H protein, human; D039642:Eukaryotic Initiation Factors; D010448:Peptide Initiation Factors; D016601:RNA-Binding Proteins