657 research outputs found

    Lrp Acts as Both a Positive and Negative Regulator for Type 1 Fimbriae Production in Salmonella enterica Serovar Typhimurium

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    Leucine-responsive regulatory protein (Lrp) is known to be an indirect activator of type 1 fimbriae synthesis in Salmonella enterica serovar Typhimurium via direct regulation of FimZ, a direct positive regulator for type 1 fimbriae production. Using RT-PCR, we have shown previously that fimA transcription is dramatically impaired in both lrp-deletion (Δlrp) and constitutive-lrp expression (lrpC) mutant strains. In this work, we used chromosomal PfimA-lacZ fusions and yeast agglutination assays to confirm and extend our previous results. Direct binding of Lrp to PfimA was shown by an electrophoretic mobility shift assay (EMSA) and DNA footprinting assay. Site-directed mutagenesis revealed that the Lrp-binding motifs in PfimA play a role in both activation and repression of type 1 fimbriae production. Overproduction of Lrp also abrogates fimZ expression. EMSA data showed that Lrp and FimZ proteins independently bind to PfimA without competitive exclusion. In addition, both Lrp and FimZ binding to PfimA caused a hyper retardation (supershift) of the DNA-protein complex compared to the shift when each protein was present alone. Nutrition-dependent cellular Lrp levels closely correlated with the amount of type 1 fimbriae production. These observations suggest that Lrp plays important roles in type 1 fimbriation by acting as both a positive and negative regulator and its effect depends, at least in part, on the cellular concentration of Lrp in response to the nutritional environment

    Gene expression profile of the skin in the 'hairpoor' (HrHp) mice by microarray analysis

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    <p>Abstract</p> <p>Background</p> <p>The transcriptional cofactor, Hairless (HR), acts as one of the key regulators of hair follicle cycling; the loss of function mutations is the cause of the expression of the hairless phenotype in humans and mice. Recently, we reported a new <it>Hr </it>mutant mouse called 'Hairpoor' (<it>Hr<sup>Hp</sup></it>). These mutants harbor a gain of the function mutation, T403A, in the <it>Hr </it>gene. This confers the overexpression of HR and <it>Hr<sup>Hp </sup></it>is an animal model of Marie Unna hereditary hypotrichosis in humans. In the present study, the expression profile of <it>Hr<sup>Hp</sup>/Hr<sup>Hp </sup></it>skin was investigated using microarray analysis to identify genes whose expression was affected by the overexpression of HR.</p> <p>Results</p> <p>From 45,282 mouse probes, differential expressions in 43 (>2-fold), 306 (>1.5-fold), and 1861 genes (>1.2-fold) in skin from <it>Hr<sup>Hp</sup>/Hr<sup>Hp </sup></it>mice were discovered and compared with skin from wild-type mice. Among the 1861 genes with a > 1.2-fold increase in expression, further analysis showed that the expression of eight genes known to have a close relationship with hair follicle development, ascertained by conducting real-time PCR on skin RNA produced during hair follicle morphogenesis (P0-P14), indicated that four genes, <it>Wif1</it>, <it>Casp14</it>, <it>Krt71</it>, and <it>Sfrp1</it>, showed a consistent expression pattern with respect to HR overexpression in vivo.</p> <p>Conclusion</p> <p><it>Wif1 </it>and <it>Casp14 </it>were found to be upregulated, whereas <it>Krt71 </it>and <it>Sfrp1 </it>were downregulated in cells overexpressing HR in transient transfection experiments on keratinocytes, suggesting that HR may transcriptionally regulate these genes. Further studies are required to understand the mechanism of this regulation by the HR cofactor.</p
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