Publications
1997
Malmgren C, Wagner E G, Ehresmann C, Ehresmann B, Romby P
Antisense RNA control of plasmid R1 replication. The dominant product of the antisense rna-mrna binding is not a full RNA duplex Journal Article
In: J Biol Chem, vol. 272, no. 19, pp. 12508-12512, 1997, ISBN: 9139701, (0021-9258 Journal Article).
Abstract | Links | BibTeX | Tags: Antisense/*metabolism RNA, Bacterial Proteins/genetics/metabolism Base Sequence Copper/metabolism Electrophoresis, Messenger/*metabolism Ribonuclease III Support, Non-U.S. Gov't, Polyacrylamide Gel Endoribonucleases/metabolism Escherichia coli Lead Molecular Sequence Data Nucleic Acid Conformation Plasmids/*metabolism Pseudomonas RNA, ROMBY, Unité ARN
@article{,
title = {Antisense RNA control of plasmid R1 replication. The dominant product of the antisense rna-mrna binding is not a full RNA duplex},
author = {C Malmgren and E G Wagner and C Ehresmann and B Ehresmann and P Romby},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9139701},
isbn = {9139701},
year = {1997},
date = {1997-01-01},
journal = {J Biol Chem},
volume = {272},
number = {19},
pages = {12508-12512},
abstract = {The replication frequency of plasmid R1 is controlled by an antisense RNA (CopA) that binds to its target site (CopT) in the leader region of repA mRNA and inhibits the synthesis of the replication initiator protein RepA. Previous studies on CopA-CopT pairing in vitro revealed the existence of a primary loop-loop interaction (kissing complex) that is subsequently converted to an almost irreversible duplex. However, the structure of more stable binding intermediates that lead to the formation of a complete duplex was speculative. Here, we investigated the interaction between CopA and CopT by using Pb(II)-induced cleavages. The kissing complex was studied using a truncated antisense RNA (CopI) that is unable to form a full duplex with CopT. Furthermore, RNase III, which is known to process the CopA-CopT complex in vivo, was used to detect the existence of a full duplex. Our data indicate that the formation of a full CopA-CopT duplex appears to be a very slow process in vitro. Unexpectedly, we found that the loop-loop interaction persists in the predominant CopA-CopT complex and is stabilized by intermolecular base pairing involving the 5'-proximal 30 nucleotides of CopA and the complementary region of CopT. This almost irreversible complex suffices to inhibit ribosome binding at the tap ribosome binding site and may be the inhibitory complex in vivo.},
note = {0021-9258
Journal Article},
keywords = {Antisense/*metabolism RNA, Bacterial Proteins/genetics/metabolism Base Sequence Copper/metabolism Electrophoresis, Messenger/*metabolism Ribonuclease III Support, Non-U.S. Gov't, Polyacrylamide Gel Endoribonucleases/metabolism Escherichia coli Lead Molecular Sequence Data Nucleic Acid Conformation Plasmids/*metabolism Pseudomonas RNA, ROMBY, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
The replication frequency of plasmid R1 is controlled by an antisense RNA (CopA) that binds to its target site (CopT) in the leader region of repA mRNA and inhibits the synthesis of the replication initiator protein RepA. Previous studies on CopA-CopT pairing in vitro revealed the existence of a primary loop-loop interaction (kissing complex) that is subsequently converted to an almost irreversible duplex. However, the structure of more stable binding intermediates that lead to the formation of a complete duplex was speculative. Here, we investigated the interaction between CopA and CopT by using Pb(II)-induced cleavages. The kissing complex was studied using a truncated antisense RNA (CopI) that is unable to form a full duplex with CopT. Furthermore, RNase III, which is known to process the CopA-CopT complex in vivo, was used to detect the existence of a full duplex. Our data indicate that the formation of a full CopA-CopT duplex appears to be a very slow process in vitro. Unexpectedly, we found that the loop-loop interaction persists in the predominant CopA-CopT complex and is stabilized by intermolecular base pairing involving the 5'-proximal 30 nucleotides of CopA and the complementary region of CopT. This almost irreversible complex suffices to inhibit ribosome binding at the tap ribosome binding site and may be the inhibitory complex in vivo.