DNA uptake sequences are widespread throughout the Neisseria gonorrhoeae genome. These short, conserved sequences facilitate the exchange of endogenous DNA between members of the genus Neisseria. Often the DNA uptake sequences are present as inverted repeats that are able to form hairpin structures. It has been suggested previously that DNA uptake sequence inverted repeats present 3 ¢ of genes play a role in rho-independent termination and attenuation. However, there is conflicting experimental evidence to support this role. The aim of this study was to determine the role of DNA uptake sequences in transcriptional termination. Both bioinformatics predictions, conducted using TransTermHP, and experimental evidence, from RNA-seq data, were used to determine which inverted repeat DNA uptake sequences are transcriptional terminators and in which direction. Here we show that DNA uptake sequences in the inverted repeat configuration occur in N. gonorrhoeae both where the DNA uptake sequence precedes the inverted version of the sequence and also, albeit less frequently, in reverse order. Due to their symmetrical configuration, inverted repeat DNA uptake sequences can potentially act as bi-directional terminators, therefore affecting transcription on both DNA strands. This work also provides evidence that gaps in DNA uptake sequence density in the gonococcal genome coincide with areas of DNA that are foreign in origin, such as prophage. This study differentiates for the first time, to our knowledge, between DNA uptake sequences that form intrinsic transcriptional terminators and those that do not, providing characteristic features within the flanking inverted repeat that can be identified.Keywords: Gonococcal; DUS; rho-independent transcriptional termination; HGT.Abbreviations: DUS, DNA uptake sequence; IR-DUS, inverted repeat DNA uptake sequence; eDUS, extended 12 bp DNA uptake sequence; vDUS, variant DNA uptake sequence found in commensal non-pathogenic Neisseria spp.; veDUS, extended 12 bp variant DNA uptake sequence; IR-eDUS, inverted repeat extended DNA uptake sequence; IR-vDUS, inverted repeat variant DNA uptake sequence; CREE, Correia repeat enclosed elements; GGI, Gonococcal genetic island.
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