Effect of DNA bending on transcriptional interference in the systems of closely spaced convergent promoters

Authors：Koroleva Olga N^*; Dubrovin Evgeniy V^*; Yaminsky Igor V; Drutsa Valeriy L

Source：BBA - General Subjects, 2016, 1860(10): 2086-2096.

DOI：10.1016/j.bbagen.2016.06.026

• Convergent transcription
• Transcription interference
• E. coli RNA polymerase
• DNA structure
• Open promoter complexes
• Atomic force microscopy

Background: Over the past years there are increasing evidences that the interplay between two molecules of RNA polymerases, initiating transcription from promoters, oriented in opposite (convergent) directions, can serve as a regulatory factor of gene expression. The data concerning the molecular mechanisms of this so-called transcriptional interference (TI) are not well understood.
Methods: The interaction of RNA polymerase with circular DNA templates, containing the convergent promoters, was investigated in a series of in vitro transcription assays and atomic force microscopy (AFM).
Results: In this work, to study the mechanisms of transcription interference a series of plasmids with oppositely oriented closely spaced artificial promoters, recognized by Escherichia coli RNA polymerase, was constructed. The constructs differ in promoter structure and distance between the transcription start sites. We have demonstrated that the transcripts ratio (RNA-R/RNA-L) and morphology of convergent open promoter complexes (OPC) are highly dependent on the interpromoter distance.
Conclusions: The obtained results allowed us to suggest the novel model of TI, which assumes the DNA bending upon binding of RNA polymerase with promoters and explains the phenomenon of complete inactivation of weaker promoter by the stronger one.
General significance: The results show that the conformational transitions in DNA helix, associated with DNA bending upon binding of RNA polymerase with promoters, play crucial role in OPC formation in the systems with convergent promoteri.

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