Alexandra Moores’s research while affiliated with University of Kent and other places

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Publications (2)


Supplemental material
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January 2014

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14 Reads

Alexandra Moores

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Lei Sun

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The effects of ΔrfaH and ΔslyA mutations on the β-galactosidase produced by FimB-LacZ translational (tl) and fimB-lacZ transcriptional (tc) fusions. The wild-type (wt) and mutant strains indicated were grown and processed as described in Materials and Methods.
The effects of ΔrfaH on FimB off-to-on recombination per cell per generation. The bacteria were grown and processed as described in Materials and Methods. The values shown are the means of at least five measurements.
The organization of the fimB promoter and 5′ UTR. The extents of deletion mutations Δ1 to Δ3 are indicated by solid lines. Also indicated are the positions of the MicA target sequence in the fimB mRNA (26), the predicted fimB Shine-Dalgarno sequence (SD), and the ops site-like element OLE. The fimB promoter −35 and −10 regions (shaded rectangles), transcriptional start site and direction (arrow), and previously characterized SlyA binding sites OSA1 and OSA2 (34) are also shown. The start of the fimB ORF is indicated by the labeled box. Sp (SphI) and EO (EcoO109I) correspond to the restriction endonuclease sites used in this study. The ClaI site used lies within the fimB ORF further downstream than the region included in the diagram. The scale of the diagram (100 bp) is indicated by an additional horizontal line. The parallel diagonal lines denote that OSA1 and OSA2 lie further upstream of the fimB promoter than indicated by the linear scale of the diagram.
The effects of Δ1 to Δ3 mutations on the β-galactosidase produced by a FimB-LacZ fusion. The wild-type (wt) and mutant strains indicated were grown and processed as described in Materials and Methods.
The effects of micA, rfaH, and micA rfaH double mutations on the β-galactosidase produced by a FimB-LacZ fusion. The wild-type (wt) and mutant strains indicated were grown and processed as described in Materials and Methods, except that the growth medium used contained 1 mM nicotinic acid to allow growth of the rseA mutants which contain a linked nadB::Tn10 mutation.

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RfaH Suppresses Small RNA MicA Inhibition of fimB Expression in Escherichia coli K-12

January 2014

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391 Reads

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6 Citations

The phase variation (reversible on-off switching) of the type 1 fimbrial adhesin of Escherichia coli involves a DNA inversion catalyzed by FimB (switching in either direction) or FimE (on-to-off switching). Here, we demonstrate that RfaH activates expression of a FimB-LacZ protein fusion while having a modest inhibitory effect on a comparable fimB-lacZ operon construct and on a FimE-LacZ protein fusion, indicating that RfaH selectively controls fimB expression at the posttranscriptional level. Further work demonstrates that loss of RfaH enables small RNA (sRNA) MicA inhibition of fimB expression even in the absence of exogenous inducing stress. This effect is explained by induction of σE, and hence MicA, in the absence of RfaH. Additional work confirms that the procaine-dependent induction of micA requires OmpR, as reported previously (A. Coornaert et al., Mol. Microbiol. 76:467–479, 2010, doi:10.1111/j.1365-2958.2010.07115.x), but also demonstrates that RfaH inhibition of fimB transcription is enhanced by procaine independently of OmpR. While the effect of procaine on fimB transcription is shown to be independent of RcsB, it was found to require SlyA, another known regulator of fimB transcription. These results demonstrate a complex role for RfaH as a regulator of fimB expression.

Citations (1)


... The Escherichia coli micA gene was the most cited bacterial antisense gene with 25 related articles in PubMed (Table S7). This gene stands out for being a post-transcriptional regulator of several genes [140][141][142] and for acting in the mechanisms of virulence [143]. A vaccine produced with micA-derived OMVs (outer membrane vesicles) protected mice against Salmonella typhimurium [143]. ...

Reference:

Antisense Transcription in Plants: A Systematic Review and an Update on cis-NATs of Sugarcane
RfaH Suppresses Small RNA MicA Inhibition of fimB Expression in Escherichia coli K-12