Intricate interactions within the ccd plasmid addiction system

Anne Daineffe; Daniel Charlier; Remy Loris; Dominique Maes; Joris Messens; Lode Wyns; Jan Backmann

Intricate interactions within the ccd plasmid addiction system

Anne Daineffe, Daniel Charlier, Remy Loris, Dominique Maes, Joris Messens, Lode Wyns, Jan Backmann

Research output: Contribution to journal › Article › peer-review

67 Citations (Scopus)

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.

Original language	English
Pages (from-to)	3733-3742
Number of pages	10
Journal	J. Biol. Chem.
Volume	277
Issue number	5
Publication status	Published - 1 Feb 2002

Keywords

killer protein
gyrase

7 Talk or presentation at a workshop/seminar
4 Participation in conference
3 Participation in workshop, seminar
3 Talk or presentation at a conference

VIB Group Leader meeting
Remy Loris (Keynote speaker)
27 Jan 2012
Activity: Talk or presentation › Talk or presentation at a workshop/seminar
International Union of Crystallography (IUCr) Congress and general Assembly 2011
Remy Loris (Keynote speaker)
22 Aug 2011 → 30 Aug 2011
Activity: Talk or presentation › Talk or presentation at a conference
IRB BioMed Conference on "Intrinsically disordered proteins in biomedicine"
Nico Van Nuland (Participant)
4 Oct 2010 → 6 Oct 2010
Activity: Participating in or organising an event › Participation in workshop, seminar

Cite this

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title = "Intricate interactions within the ccd plasmid addiction system",

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.",

keywords = "killer protein, gyrase",

author = "Anne Daineffe and Daniel Charlier and Remy Loris and Dominique Maes and Joris Messens and Lode Wyns and Jan Backmann",

year = "2002",

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language = "English",

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pages = "3733--3742",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

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TY - JOUR

T1 - Intricate interactions within the ccd plasmid addiction system

AU - Daineffe, Anne

AU - Charlier, Daniel

AU - Loris, Remy

AU - Maes, Dominique

AU - Messens, Joris

AU - Wyns, Lode

AU - Backmann, Jan

PY - 2002/2/1

Y1 - 2002/2/1

N2 - 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.

AB - 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.

KW - killer protein

KW - gyrase

M3 - Article

VL - 277

SP - 3733

EP - 3742

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 5

ER -

Intricate interactions within the ccd plasmid addiction system

Abstract

Keywords

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Activities

VIB Group Leader meeting

International Union of Crystallography (IUCr) Congress and general Assembly 2011

IRB BioMed Conference on "Intrinsically disordered proteins in biomedicine"

Cite this