Nucleolytic inactivation and degradation of the RNase III processed pnp message encoding polynucleotide phosphorylase of Escherichia coli.
The two cleavages made by RNase III in the transcripts of the pnp gene of Escherichia coli, 80 nucleotides upstream of the coding sequence of polynucleotide phosphorylase, were previously demonstrated to trigger the rapid degradation of the pnp messenger. In this paper, we demonstrate that the 5' end of the RNase III processed pnp mRNA is attacked by ribonucleases more efficiently than the rest of the molecule. Several 5' extremities resulting from cleavages occurring in the first 500 nucleotides of the pnp transcript have been identified. Three of them referred to as X, Y and W occur in the wild-type strain at the beginning of the coding sequence of the pnp mRNA. The mRNA appears to be cleaved more efficiently at the X site, proximal to the initiation codon, than at sites Y and W located downstream. In vitro, the maturation at X is catalysed by RNase E but not by RNase III. Accumulation of RNA processed at X in RNase E deficient strains leads us to postulate that X is a high affinity primary site which is slowly cleaved by the residual activity of thermosensitive RNase E at non-permissive temperature and that secondary sites located downstream are processed less efficiently than X. Taken together, our results suggest that in wild-type E. coli the degradation of the RNase III processed mRNA is mediated by RNase E.
Hajnsdorf
E
E
CNRS URA1139, Institut de Biologie Physico-chimique, Paris, France.
Carpousis
A J
AJ
Régnier
P
P
D001483:Base Sequence; D001665:Binding Sites; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D011117:Polyribonucleotide Nucleotidyltransferase / Q000235:genetics*; D015347:RNA Probes / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional / Q000502:physiology*; D012329:RNA, Bacterial / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D043244:Ribonuclease III; D013696:Temperature; D014158:Transcription, Genetic / Q000235:genetics
D029968:Escherichia coli Proteins; D015347:RNA Probes; D012329:RNA, Bacterial; D012333:RNA, Messenger; D011117:Polyribonucleotide Nucleotidyltransferase; D004722:Endoribonucleases; D043244:Ribonuclease III; C468366:ribonuclease III, E coli; C022537:ribonuclease E