PMID:12206755
| Citation |
Xu, FF, Gaggero, C and Cohen, SN (2002) Polyadenylation can regulate ColE1 type plasmid copy number independently of any effect on RNAI decay by decreasing the interaction of antisense RNAI with its RNAII target. Plasmid 48:49-58 |
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| Abstract |
Replication of ColE1-type plasmids is regulated by RNAI, an antisense RNA that interacts with the replication pre-primer, RNAII. Exonucleolytic attack at the 3' end of RNAI is impeded in pcnB mutant bacteria, which lack poly(A) polymerase I-the principal RNA polyadenylase of E. coli; this leads to accumulation of an RNAI decay intermediate (RNAI(-5)) and dramatic reduction of the plasmid copy number. Here, we report that polyadenylation can also affect RNAI-mediated control of plasmid DNA replication by inhibiting interaction of RNAI(-5) with RNAII. We show that mutation of the host pcnB gene profoundly affects the plasmid copy number, even under experimental conditions that limit the effects of polyadenylation on RNAI(-5) decay. Moreover, poly(A) tails interfere with RNAI/RNAII interaction in vitro without producing any detectable alteration of RNAI secondary structure. Our results establish the existence of a previously undetected mechanism by which RNA polyadenylation can control plasmid copy number. |
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| Keywords |
Bacterial Proteins/genetics; Base Sequence; DNA Primers; Escherichia coli/genetics; Gene Expression Regulation, Bacterial; Molecular Sequence Data; Nucleic Acid Conformation; Plasmids/genetics; Poly A/genetics; Polymerase Chain Reaction; RNA, Antisense/genetics; RNA, Antisense/metabolism; RNA, Bacterial/genetics; RNA, Small Interfering |
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