Intricate interactions within the ccd plasmid addiction system.

TitleIntricate interactions within the ccd plasmid addiction system.
Publication TypeJournal Article
Year of Publication2002
AuthorsDao-Thi, M-H., D. Charlier, R. Loris, D. Maes, J. Messens, L. Wyns, and J. Backmann
JournalJ Biol Chem
Volume277
Issue5
Pagination3733-42
Date Published2002 Feb 1
ISSN0021-9258
KeywordsBacterial Proteins, Bacterial Toxins, Cytotoxins, Escherichia coli, F Factor, Kinetics, Models, Molecular, Protein Conformation, Spectrophotometry, Ultraviolet
Abstract

The ccd addiction system plays a crucial role in the stable maintenance of the Escherichia coli F plasmid. It codes for a stable toxin (CcdB) and a less stable antidote (CcdA). Both are expressed at low levels during normal cell growth. Upon plasmid loss, CcdB outlives CcdA and kills the cell by poisoning gyrase. The interactions between CcdB, CcdA, and its promoter DNA were analyzed. In solution, the CcdA-CcdB interaction is complex, leading to various complexes with different stoichiometry. CcdA has two binding sites for CcdB and vice versa, permitting soluble hexamer formation but also causing precipitation, especially at CcdA:CcdB ratios close to one. CcdA alone, but not CcdB, binds to promoter DNA with high on and off rates. The presence of CcdB enhances the affinity and the specificity of CcdA-DNA binding and results in a stable CcdA*CcdB*DNA complex with a CcdA:CcdB ratio of one. This (CcdA(2)CcdB(2))(n) complex has multiple DNA-binding sites and spirals around the 120-bp promoter region.

DOI10.1074/jbc.M105505200
Alternate JournalJ. Biol. Chem.
PubMed ID11741897
subject_category: 
Research group: