Identification and characterization of DUSP27, a novel dual-specific protein phosphatase

AbstractA novel human dual-specific protein phosphatase (DSP), designated DUSP27, is here described. The DUSP27 gene contains three exons, rather than the predicted 4–14 exons, and encodes a 220 amino acid protein. DUSP27 is structurally similar to other small DSPs, like VHR and DUSP13. The location of DUSP27 on chromosome 10q22, 50kb upstream of DUSP13, suggests that these two genes arose by gene duplication. DUSP27 is an active enzyme, and its kinetic parameters and were determined. DUSP27 is a cytosolic enzyme, expressed in skeletal muscle, liver and adipose tissue, suggesting its possible role in energy metabolism

Ler Is a Negative Autoregulator of the LEE1 Operon in Enteropathogenic Escherichia coli

Enteropathogenic Escherichia coli (EPEC) causes severe diarrhea in young children. Essential for colonization of the host intestine is the LEE pathogenicity island, which comprises a cluster of operons encoding a type III secretion system and related proteins. The LEE1 operon encodes Ler, which positively regulates many EPEC virulence genes in the LEE region and elsewhere in the chromosome. We found that Ler acts as a specific autorepressor of LEE1 transcription. We further show that Ler specifically binds upstream of the LEE1 operon in vivo and in vitro. A comparison of the Ler affinities to different DNA regions suggests that the autoregulation mechanism limits the steady-state level of Ler to concentrations that are just sufficient for activation of the LEE2 and LEE3 promoters and probably other LEE promoters. This mechanism may reflect the need of EPEC to balance maximizing the colonization efficiency by increasing the expression of the virulence genes and minimizing the immune response of the host by limiting their expression. In addition, we found that the autoregulation mechanism reduces the cell-to-cell variability in the levels of LEE1 expression. Our findings point to a new negative regulatory circuit that suppresses the noise and optimizes the expression levels of ler and other LEE1 genes

Effects of the Medium Composition on the Components of the Electrochemical Proton Gradient in Rhodopseudomonas sphaeroides

The magnitude and composition of the proton motive force (Δµ~H+) has been measured in chromatophores and whole cells of Rhodopseudomonas sphaeroides as a function of the ionic composition of the buffer in which the energy-transducing membranes are suspended. Measurements with the flow-dialysis technique are compared with spectrophotometric measurements of the absorbance changes of carotenoids and of the quenching of the fluorescence of 9-aminoacridine. It is concluded that the optical techniques give rise to an overestimation of the gradients of pH and electrical potential. However, with the optical techniques the relative permeability of the membrane of R. sphaeroides for various cations and anions can be estimated. The results indicate that in general anions are more permeant than cations; the relative permeability of the anions is: SO4= < HPO4= < Cl- < NO3- < ClO4-

Energy Coupling of Facilitated Transport of Inorganic Ions in Rhodopseudomonas sphaeroides

Within the scope of a study on the effects of changes in medium composition on the proton motive force in Rhodopseudomonas sphaeroides, the energy coupling of sodium, phosphate, and potassium (rubidium) transport was investigated. Sodium was transported via an electroneutral exchange system against protons. The system functioned optimally at pH 8 and was inactive below pH 7. The driving force for the phosphate transport varied with the external pH. At pH 8, Pi transport was dependent exclusively on Δψ (transmembrane electrical potential), whereas at pH 6 only the ΔpH (transmembrane pH gradient) component of the proton motive force was a driving force. Potassium (rubidium) transport was facilitated by a transport system which catalyzed the electrogenic transfer of potassium (rubidium) ions. However, in several aspects the properties of this transport system were different from those of a simple electrogenic potassium ionophore such as valinomycin: (i) accumulated potassium leaked very slowly out of cells in the dark; and (ii) the transport system displayed a threshold in the Δψ, below which potassium (rubidium) transport did not occur