Effect of a Purified Initiation Factor F3 (B) on the Selection of Ribosomal Binding Sites on Phage MS2 RNA

Abstract

STUDIES with T4 mRNA showed that initiation factor F2 (C) promotes the attachment of ribosomes to mRNA¹. On the 30S ribosomal subunit this effect is independent of the function of F2 in the binding of formylmethionyl tRNA², whereas formation of a 70S-mRNA complex depends on the binding of fMet-tRNA³. Template competition experiments⁴ showed that, with F2 (C), the ribosome seems to have the same affinity for synthetic polynucleotides as for natural mRNA. Addition of initiation factor F3 (B), however, leads to preferential binding of ribosomes to the natural mRNA. This suggests⁴ that while factor F2 (C) binds the ribosome to any site on the mRNA, the function of factor F3 (B) is to recognize some specific signal in natural mRNA corresponding, perhaps, to the beginning of a cistron. Fractionation of initiation factor F3 (B) into several species differing in their specificity for different mRNA templates⁵ gave further support to the hypothesis that this protein can select binding sites. An excellent system to demonstrate this effect of F3 (B) would be the binding of ribosomes to RNA from E. coli RNA bacteriophages, since Steitz⁶ has analysed and determined the nucleotide sequence of the three binding sites corresponding to the three cistrons of R17 mRNA. Experiments were thus undertaken to study the effect of a purified fraction of F3 (B) on the binding of ribosomes to the different sites of such a phage RNA.