Regulation of plasmid R1 replication: PcnB and RNase E expedite the decay of the antisense RNA, CopA.
The replication frequency of plasmid R1 is controlled by an unstable antisense RNA, CopA, which, by binding to its complementary target, blocks translation of the replication rate-limiting protein RepA. Since the degree of inhibition is directly correlated with the intracellular concentration of CopA, factors affecting CopA turnover can also alter plasmid copy number. We show here that PcnB (PAPl-a poly(A)polymerase of Escherichia coli) is such a factor. Previous studies have shown that the copy number of ColE1 is decreased in pcnB mutant strains because the stability of the RNase E processed form of RNAI, the antisense RNA regulator of ColE1 replication, is increased. We find that, analogously, the twofold reduction in R1 copy number caused by a pcnB lesion is associated with a corresponding increase in the stability of the RNase E-generated 3' cleavage product of CopA. These results suggest that CopA decay is initiated by RNase E cleavage and that PcnB is involved in the subsequent rapid decay of the 3' CopA stem-loop segment. We also find that, as predicted, under conditions in which CopA synthesis is unaffected, pcnB mutation reduces RepA translation and increases CopA stability to the same extent.
Söderbom
F
F
Department of Microbiology, Biomedical Center, Uppsala University, Sweden.
Binnie
U
U
Masters
M
M
Wagner
E G
EG
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002874:Chromosome Mapping; D004261:DNA Replication; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins; D018628:Gene Dosage; D009154:Mutation; D009690:Nucleic Acid Conformation; D011062:Polynucleotide Adenylyltransferase; D011815:R Factors / Q000235:genetics*; D016372:RNA, Antisense / Q000378:metabolism*; D012329:RNA, Bacterial / Q000378:metabolism
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D016372:RNA, Antisense; D012329:RNA, Bacterial; D011062:Polynucleotide Adenylyltransferase; C063643:pcnB protein, E coli; D004722:Endoribonucleases; C022537:ribonuclease E