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Leucine-Responsive Regulatory Protein Lrp and PapI Homologues Influence Phase Variation of CS31A Fimbriae

Author:
Graveline, Richard, Garneau, Philippe, Martin, Christine, Mourez, Michaël, Hancock, Mark A., Lavoie, Rémi, Harel, Josée
Source:
Journal of bacteriology 2014 v.196 no.16 pp. 2944-2953
ISSN:
0021-9193
Subject:
antigens, bacteriology, deoxyribonuclease I, enterotoxigenic Escherichia coli, fimbriae, operon, surface plasmon resonance, transcription (genetics)
Abstract:
CS31A, a K88-related surface antigen specified by the clp operon, is a member of the type P family of adhesive factors and plays a key role in the establishment of disease caused by septicemic and enterotoxigenic Escherichia coli strains. Its expression is under the control of methylation-dependent transcriptional regulation, for which the leucine-responsive regulatory protein (Lrp) is essential. CS31A is preferentially in the OFF state and exhibits distinct regulatory features compared to the regulation of other P family members. In the present study, surface plasmon resonance and DNase I protection assays showed that Lrp binds to the distal moiety of the clp regulatory region with low micromolar affinity compared to its binding to the proximal moiety, which exhibits stronger, nanomolar affinity. The complex formation was also influenced by the addition of PapI or FooI, which increased the affinity of Lrp for the clp distal and proximal regions and was required to induce phase variation. The influence of PapI or FooI, however, was predominantly associated with a more complete shutdown of clp expression, in contrast to what has previously been observed with AfaF (a PapI ortholog). Taken together, these results suggest that the preferential OFF state observed in CS31A cells is mainly due to the weak interaction of the leucine-responsive regulatory protein with the clp distal region and that the PapI homolog favors the OFF phase. Within the large repertoire of fimbrial variants in the P family, our study illustrates that having a fimbrial operon that lacks its own PapI ortholog allows it to be more flexibly regulated by other orthologs in the cell.
Agid:
1184288