We have studied the interactions between the three molecules Rop, RNAI and RNAII that are involved in the regulatory mechanism controlling the replication of ColE1 plasmids. We show that it is possible to purify the two RNA molecules by passing an RNA mixture through an affinity column containing Rop immobilized to a solid support. The dissociation constants of the Rop-RNAI and Rop-RNAII complexes are of the order of 10(-4) M, several orders of magnitude higher than dissociation constants of stable protein-nucleic acid complexes (10(-10) M in the lambda repressor system). Although complete RNAI molecules have higher affinity, stem-and-loop I alone can also bind Rop, suggesting that this structure plays an important role in the interaction. Rop protects the stems of RNAI and RNAII from digestion by RNases while the sensitivity of the loops to digestion by RNase T1 is not affected by high concentrations of Rop. We propose a model for Rop-RNAI/RNAII interaction in which the dimeric protein acts as an adaptor between stem structures to position the two RNAs in the correct position for loop interaction.

HELMER CITTERICH, M., Anceschi, M., Banner, D., Cesareni, G. (1988). Control of ColE1 replication: low affinity specific binding of Rop (Rom) to RNAI and RNAII. EMBO JOURNAL, 7(2), 557-566.

Control of ColE1 replication: low affinity specific binding of Rop (Rom) to RNAI and RNAII

HELMER CITTERICH, MANUELA;CESARENI, GIOVANNI
1988-02-01

Abstract

We have studied the interactions between the three molecules Rop, RNAI and RNAII that are involved in the regulatory mechanism controlling the replication of ColE1 plasmids. We show that it is possible to purify the two RNA molecules by passing an RNA mixture through an affinity column containing Rop immobilized to a solid support. The dissociation constants of the Rop-RNAI and Rop-RNAII complexes are of the order of 10(-4) M, several orders of magnitude higher than dissociation constants of stable protein-nucleic acid complexes (10(-10) M in the lambda repressor system). Although complete RNAI molecules have higher affinity, stem-and-loop I alone can also bind Rop, suggesting that this structure plays an important role in the interaction. Rop protects the stems of RNAI and RNAII from digestion by RNases while the sensitivity of the loops to digestion by RNase T1 is not affected by high concentrations of Rop. We propose a model for Rop-RNAI/RNAII interaction in which the dimeric protein acts as an adaptor between stem structures to position the two RNAs in the correct position for loop interaction.
1-feb-1988
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore BIO/11 - BIOLOGIA MOLECOLARE
Settore BIO/18 - GENETICA
English
Con Impact Factor ISI
Coliphages; RNA, Bacterial; DNA Restriction Enzymes; Escherichia coli; Bacteriocin Plasmids; Nucleic Acid Conformation; Base Sequence; DNA Replication; Molecular Sequence Data; Plasmids; Transcription, Genetic
HELMER CITTERICH, M., Anceschi, M., Banner, D., Cesareni, G. (1988). Control of ColE1 replication: low affinity specific binding of Rop (Rom) to RNAI and RNAII. EMBO JOURNAL, 7(2), 557-566.
HELMER CITTERICH, M; Anceschi, M; Banner, D; Cesareni, G
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/15498
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