Mutations in PurBox1 of theBacillus subtilis pur operon control site affect adenine-regulated expressionin vivo

Transcription of theBacillus subtilis pur operon is regulated by a purine repressor (PurR)-DNA control site interaction. Thepur operon control site has two PurBoxes that are required for high-affinity PurR binding. An upstream, strong-binding PurBox1 is at position −81 to −68 relative to the transcr...

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Bibliographic Details
Published in:Science China. Life sciences 2005-09, Vol.48 (2), p.133-138
Main Authors: Xuan, Jinsong, Zalkin, Howard, Weng, Manli
Format: Article
Language:English
Subjects:
Mutation
Titration
Transcription initiation
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Summary:Transcription of theBacillus subtilis pur operon is regulated by a purine repressor (PurR)-DNA control site interaction. Thepur operon control site has two PurBoxes that are required for high-affinity PurR binding. An upstream, strong-binding PurBox1 is at position −81 to −68 relative to the transcription start site and a downstream weak-binding PurBox2 is at position −49 to −36. We constructed three PurBox1 mutations and the effects on binding of PurR to the control regionin vitro and on regulation ofpur operon expressionin vivo were investigated. The mutations significantly reduced the binding of PurR to control region DNA. In strains with G-75A, G-75T and a five bp deletion (Δ5)pur operon repression was defectivein vivo. In additionin vivo PurR titration was used to confirm that sequences flanking PurBox1 and PurBox2 are required for PurR binding to thepur operon control site.
ISSN:1006-9305
1674-7305
1862-2798
1869-1889
DOI:10.1007/BF02879665