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Volumn 250, Issue 4980, 1990, Pages 528-532

DNA looping and unlooping by AraC protein

Author keywords

[No Author keywords available]

Indexed keywords

ESCHERICHIA COLI;

EID: 0025202249     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.2237403     Document Type: Article
Times cited : (190)

References (50)
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    • in preparation
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    • Lobell, R.1    Schleif, R.2
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    • note
    • 2 and araI. A stable complex, which dissociates with a half-time of 100 minutes at 100 mM KCl (Fig. 2, wild type), is formed at low AraC concentrations in the absence of competitor DNA. The second complex is unstable, migrates slightly faster than the stable complex (Fig. 2, araI mutant, at 0 and 8 minutes) and is formed only at high AraC concentrations. Weakly associated AraC dissociates from the unstable complex with a half-time of less than 1 minute at 100 mM KCl, and the process, the stable complex is formed. In our other experiments binding reactions are incubated with an excess of unlabeled competitor DNA containing AraC binding sites so that only the stable complex is present.
  • 33
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    • note
    • 2.
  • 34
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    • note
    • 2.
  • 35
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    • note
    • On the basis of their DNase I protection experiments, Lee et al. (10) suggested that twice as much AraC might bind to araI in the presence of arabinose than in its absence. Three lines of evidence argue against this idea. (i) The retardation on electrophoresis of an araI-containing DNA fragment on the binding of AraC protein in the presence or absence of arabinose is the same, suggesting that the same amount of AraC binds in either case. In view of previous results (17), this is likely a dimer. (ii) Using the methods described above, we find that the same amount of AraC binds to araI on linear DNA in the presence and absence of arabinose (24). (iii) When linear DNA containing an AraC-arabinose-araI complex is incubated with less than stoichiometric amounts of an AraC monoclonal antibody, a single antibody-AraC-araI complex is observed on electrophoresis (24). If a dimer of AraC were to bind to each half of araI, two different antibody-AraC-araI complexes should have been observed; one complex with one antibody molecule, and one with two.
  • 41
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    • unpublished data
    • 2 also weakens the looped complex (R. Lobell, unpublished data).
    • Lobell, R.1
  • 44
    • 85033610943 scopus 로고
    • thesis, Brandeis University
    • R. Lobell, thesis, Brandeis University (1990).
    • (1990)
    • Lobell, R.1
  • 45
    • 85033615227 scopus 로고    scopus 로고
    • unpublished data
    • 2 mutant in place of the wild-type araI site (R. Lobell, unpublished data).
    • Lobell, R.1
  • 50
    • 85033616095 scopus 로고    scopus 로고
    • note
    • 2 mutation. Supported by NIH grant GM18277.


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