Non-canonical Structures in Promoter Modulate Gene Expression in Escherichia coli

Abstract

Herein we show how sequences that can form different non-canonical structures affect gene expression levels when inserted in the core of σ 70-dependent promoter, between the −35 and −10 elements recognized by RNA polymerase, in E. coli. We note that influence on level of GFP expression varies considerably depending on introduction of non-canonical structural elements in the antisense and sense strands as well as with their propensities to form G-triplex, G-hairpin, hairpin or G-quadruplex structures. Moreover, the extent of repression of expression does not relate to the in vitro thermal stability in a simple manner. Repression is most likely caused by steric interference rather than improper distance between the −35 and −10 elements. Although properties like thermal stability and topology can be somewhat different under in vivo and in vitro conditions, our results suggest that the extent of expression suppression cannot be dependent solely on thermal stabilities of Grich structures alone.

Oxidative Modification of Guanine in a Potential Z-DNA-Forming Sequence of a Gene Promoter Impacts Gene Expression

“…Mounting evidence supports formation of G4s, 33,34 iMs, 35,36 cruciform DNA, 37 triplexes, 38 and Z-DNA 38,39 in human genomic DNA, and examples exist supporting the idea that these non-canonical structures can regulate transcription in human cells, 34,38,40,41 and prokaryotic cells. 42,43 An open question remaining is whether non-canonical structures beyond G4s, 12,13 iMs, 27 and hairpins, 20 once oxidatively modified at G, can regulate mRNA expression during oxidative stress conditions. Regions with the potential to adopt Z-DNA are G rich, and should be susceptible to oxidation.…”