Tags

Type your tag names separated by a space and hit enter

Control of ColE1 replication: low affinity specific binding of Rop (Rom) to RNAI and RNAII.
EMBO J 1988; 7(2):557-66EJ

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.

Authors+Show Affiliations

European Molecular Biology Laboratory, Heidelberg, FRG.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

2452732

Citation

Helmer-Citterich, M, et al. "Control of ColE1 Replication: Low Affinity Specific Binding of Rop (Rom) to RNAI and RNAII." The EMBO Journal, vol. 7, no. 2, 1988, pp. 557-66.
Helmer-Citterich M, Anceschi MM, Banner DW, et al. Control of ColE1 replication: low affinity specific binding of Rop (Rom) to RNAI and RNAII. EMBO J. 1988;7(2):557-66.
Helmer-Citterich, M., Anceschi, M. M., Banner, D. W., & Cesareni, G. (1988). Control of ColE1 replication: low affinity specific binding of Rop (Rom) to RNAI and RNAII. The EMBO Journal, 7(2), pp. 557-66.
Helmer-Citterich M, et al. Control of ColE1 Replication: Low Affinity Specific Binding of Rop (Rom) to RNAI and RNAII. EMBO J. 1988;7(2):557-66. PubMed PMID: 2452732.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Control of ColE1 replication: low affinity specific binding of Rop (Rom) to RNAI and RNAII. AU - Helmer-Citterich,M, AU - Anceschi,M M, AU - Banner,D W, AU - Cesareni,G, PY - 1988/2/1/pubmed PY - 1988/2/1/medline PY - 1988/2/1/entrez SP - 557 EP - 66 JF - The EMBO journal JO - EMBO J. VL - 7 IS - 2 N2 - 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. SN - 0261-4189 UR - https://www.unboundmedicine.com/medline/citation/2452732/Control_of_ColE1_replication:_low_affinity_specific_binding_of_Rop__Rom__to_RNAI_and_RNAII_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0261-4189&date=1988&volume=7&issue=2&spage=557 DB - PRIME DP - Unbound Medicine ER -