A fluorescence study of the interaction of protein synthesis initiation factors 4A, 4E, and 4F with mRNA and oligonucleotide analogs.
The initial interaction of mRNA with the protein synthesis machinery presumably involves recognition of the 5'-cap (m7GpppN), although it is not clear at the present time whether this recognition is by eIF-4E or eIF-4F. This process has been studied by direct fluorescence titration experiments. The equilibrium constants for the formation of the binary protein: m7GpppG, protein:mRNA, and protein:protein complexes as well as the ternary mRNA:eIF-4E:eIF-4A complexes were measured. These studies show, for the first time, direct evidence for an eIF-4A:eIF-4E interaction. In contrast to earlier studies, we show that the affinity of eIF-4E and eIF-4F for globin mRNA is similar. Furthermore, the relative affinities of mRNA analogs (capped oligonucleotides) for these initiation factors indicate that the cap is the predominant feature recognized for binding, but other features also contribute to the eIF-4E:mRNA interaction.
Goss
D J
DJ
Department of Chemistry, Hunter College, City University of New York, NY 10021.
Carberry
S E
SE
Dever
T E
TE
Merrick
W C
WC
Rhoads
R E
RE
D000818:Animals; D001483:Base Sequence; D039601:Eukaryotic Initiation Factor-4A; D039561:Eukaryotic Initiation Factor-4E; D039562:Eukaryotic Initiation Factor-4F; D005914:Globins / Q000235:genetics; D007700:Kinetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000378:metabolism*; D010448:Peptide Initiation Factors / Q000378:metabolism*; D011485:Protein Binding; D012315:RNA Caps / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D011817:Rabbits; D013050:Spectrometry, Fluorescence
D039561:Eukaryotic Initiation Factor-4E; D039562:Eukaryotic Initiation Factor-4F; D009843:Oligoribonucleotides; D010448:Peptide Initiation Factors; D012315:RNA Caps; D012333:RNA, Messenger; D005914:Globins; D039601:Eukaryotic Initiation Factor-4A