Analyses of frameshifting at UUU-pyrimidine sites.
Others have recently shown that the UUU phenylalanine codon is highly frameshift-prone in the 3'(rightward) direction at pyrimidine 3'contexts. Here, several approaches are used to analyze frameshifting at such sites. The four permutations of the UUU/C (phenylalanine) and CGG/U (arginine) codon pairs were examined because they vary greatly in their expected frameshifting tendencies. Furthermore, these synonymous sites allow direct tests of the idea that codon usage can control frameshifting. Frameshifting was measured for these dicodons embedded within each of two broader contexts: the Escherichia coli prfB (RF2 gene) programmed frameshift site and a 'normal' message site. The principal difference between these contexts is that the programmed frameshift contains a purine-rich sequence upstream of the slippery site that can base pair with the 3'end of 16 S rRNA (the anti-Shine-Dalgarno) to enhance frameshifting. In both contexts frameshift frequencies are highest if the slippery tRNAPhe is capable of stable base pairing in the shifted reading frame. This requirement is less stringent in the RF2 context, as if the Shine-Dalgarno interaction can help stabilize a quasi-stable rephased tRNA:message complex. It was previously shown that frameshifting in RF2 occurs more frequently if the codon 3'to the slippery site is read by a rare tRNA. Consistent with that earlier work, in the RF2 context frameshifting occurs substantially more frequently if the arginine codon is CGG, which is read by a rare tRNA. In contrast, in the 'normal' context frameshifting is only slightly greater at CGG than at CGU. It is suggested that the Shine-Dalgarno-like interaction elevates frameshifting specifically during the pause prior to translation of the second codon, which makes frameshifting exquisitely sensitive to the rate of translation of that codon. In both contexts frameshifting increases in a mutant strain that fails to modify tRNA base A37, which is 3'of the anticodon. Thus, those base modifications may limit frameshifting at UUU codons. Finally, statistical analyses show that UUU Ynn dicodons are extremely rare in E.coli genes that have highly biased codon usage.
Schwartz
R
R
Department of Biology, Wake Forest University, PO Box 7325, Winston-Salem, NC 27109, USA.
Curran
J F
JF
D001120:Arginine / Q000235:genetics; D001482:Base Composition; D001483:Base Sequence; D003062:Codon; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics; D018965:Frameshifting, Ribosomal; D005821:Genetic Techniques; D010649:Phenylalanine / Q000235:genetics; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D014176:Protein Biosynthesis; D012347:RNA, Transfer, Arg / Q000235:genetics*; D012360:RNA, Transfer, Phe / Q000235:genetics*; D015183:Restriction Mapping; D012475:Salmonella / Q000235:genetics; D014498:Uracil
D003062:Codon; D017931:DNA Primers; D012347:RNA, Transfer, Arg; D012360:RNA, Transfer, Phe; D010649:Phenylalanine; D014498:Uracil; D001120:Arginine