Participation of IHF and a distant UP element in the stimulation of the phage λ PL promoter

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Abstract

Summary

We have previously identified a UP element in the phage λ PL promoter, centred at position −90 from the transcription start site. Integration host factor (IHF), a heterodimeric DNA-binding and -bending protein, binds upstream of the λ PL promoter in a region overlapping the UP element. Stimulation of transcription by IHF requires an intact αCTD and affects the initial binding of RNA polymerase to the promoter. We propose a model for the stimulation of PL by IHF in which IHF bends the DNA to bring the distal UP sequence in closer proximity to the promoter core sequences to allow the docking of the αCTD of RNA polymerase. Furthermore, IHF may also participate in protein-protein interactions with the αCTD. In support of this model, we found that alanine substitutions in αCTD at positions 265, 268, 270 and 275 reduced PL promoter activity. Mutations in the IHF DNA binding site, as well as IHF mutant proteins exhibiting a decreased ability to bend the DNA, were both defective in stimulating the PL promoter. In addition, some of the mutated IHF residues are clustered at a protein surface that interacts with the UP DNA sequence. These residues may also participate in protein-protein interactions with the αCTD.

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