Cleavage of 16S rRNA within the ribosome by mRNA modified in the A-site codon with phenanthroline-Cu(II).
Cleavage of 16S rRNA was obtained through mRNA modified at position +5 with the chemical cleavage agent 1,10-o-phenanthroline. In the presence of Cu2+, and after addition of reducing agent to the modified mRNA-70S complex, cleavage of proximal nucleotides within the 16S rRNA occurred. Primer extension analysis of 16S rRNA fragments revealed that nucleotides 528-532, 1196, and 1396-1397 were cleaved. Nucleotides 1053-1055 were also cleaved but did not show the same level of specificity as the former. These results provide evidence that at some point in the translation process these regions are all within 15 A of position +5, the A-site codon, on the mRNA.
Bucklin
D J
DJ
Division of Biological Sciences, University of Montana, Missoula, Montana 59812, USA.
van Waes
M A
MA
Bullard
J M
JM
Hill
W E
WE
D001483:Base Sequence; D001665:Binding Sites; D003062:Codon; D008969:Molecular Sequence Data; D010618:Phenanthrolines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000187:drug effects* / Q000378:metabolism*; D012336:RNA, Ribosomal, 16S / Q000378:metabolism*; D012266:Ribose / Q000378:metabolism*; D014475:Uncoupling Agents / Q000494:pharmacology*
D003062:Codon; D010618:Phenanthrolines; D012333:RNA, Messenger; D012336:RNA, Ribosomal, 16S; D014475:Uncoupling Agents; C014741:bis(1,10-phenanthroline)copper(2+) ion; D012266:Ribose