An interaction of wheat germ initiation factor 4B with oligoribonucleotides.

Abstract

The binding of oligoribonucleotides to wheat germ protein synthesis initiation factor eIF-4B was measured by direct fluorescence techniques. An analysis of the equilibrium association constants (Keq) indicates that eIF-4B binding is not affected by the m7GTP cap structure or the AUG. eIF-4B is insensitive to hairpin structures within the oligoribonucleotide. The binding site size is approximately 18 bases. The binding of oligoribonucleotide to eIF-4B as a function of pH, temperature, and ionic strength is also described. The pH-dependent binding showed an increase in binding with increasing pH in contrast to the sharp pH optimum observed for cap binding protein eIF-4E (Carberry, S. E., Darzynkiewicz, E., and Goss, D.J. (1991) Biochemistry 30, 1624-1627). Assuming all tryptophan residues contribute to the observed fluorescence, iodide quenching showed that 8(+/- 1) out 9 of eIF-4B's tryptophan residues are on the surface of the eIF-4B protein. A specific anion effect of Cl- on eIF-4B binding to oligoribonucleotide was found when comparing the ionic strength effect of KC2H3O2 and KCl.