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Pedro V Peña, Steven Glasker, Friedrich Srienc
The production of biofuels from cellulosic biomass is a promising technology for developing a renewable source of energy. Efforts to produce ethanol from cellulosic biomass using microbes, such as the yeast Saccharomyces cerevisiae, face major challenges, including the need for detoxification. Here, we apply a strategy to discover genetic alterations that lead to improved robustness of S. cerevisiae in the presence of acetate, which is present at toxic concentrations in hemicellulose hydrolysates. Acetate in its protonated form (acetic acid) enters the cell through passive diffusion and dissociates into a proton and acetate, acidifying the cytosol and inhibiting cell function, an effect that is exacerbated in the presence of sodium...
March 10, 2013: Journal of Biotechnology
Tomoko Nozoye, Seiji Nagasaka, Takanori Kobayashi, Michiko Takahashi, Yuki Sato, Yoko Sato, Nobuyuki Uozumi, Hiromi Nakanishi, Naoko K Nishizawa
Eukaryotic organisms have developed diverse mechanisms for the acquisition of iron, which is required for their survival. Graminaceous plants use a chelation strategy. They secrete phytosiderophore compounds, which solubilize iron in the soil, and then take up the resulting iron-phytosiderophore complexes. Bacteria and mammals also secrete siderophores to acquire iron. Although phytosiderophore secretion is crucial for plant growth, its molecular mechanism remains unknown. Here, we show that the efflux of deoxymugineic acid, the primary phytosiderophore from rice and barley, involves the TOM1 and HvTOM1 genes, respectively...
February 18, 2011: Journal of Biological Chemistry
Begoña Benito, Blanca Garciadeblás, José Pérez-Martín, Alonso Rodríguez-Navarro
Potassium and Na(+) effluxes across the plasma membrane are crucial processes for the ionic homeostasis of cells. In fungal cells, these effluxes are mediated by cation/H(+) antiporters and ENA ATPases. We have cloned and studied the functions of the two ENA ATPases of Ustilago maydis, U. maydis Ena1 (UmEna1) and UmEna2. UmEna1 is a typical K(+) or Na(+) efflux ATPase whose function is indispensable for growth at pH 9.0 and for even modest Na(+) or K(+) tolerances above pH 8.0. UmEna1 locates to the plasma membrane and has the characteristics of the low-Na(+)/K(+)-discrimination ENA ATPases...
June 2009: Eukaryotic Cell
Alan Gilbert, Dipen P Sangurdekar, Friedrich Srienc
Acetate is present in lignocellulosic hydrolysates at growth inhibiting concentrations. Industrial processes based on such feedstock require strains that are tolerant of this and other inhibitors present. We investigated the effect of acetate on Saccharomyces cerevisiae and show that elevated acetate concentrations result in a decreased specific growth rate, an accumulation of cells in the G1 phase of the cell cycle, and an increased cell size. With the cytostat cultivation technology under previously derived optimal operating conditions, several acetate resistant mutants were enriched and isolated in the shortest possible time...
June 15, 2009: Biotechnology and Bioengineering
Yasuo Watanabe, Yasutaka Shimono, Hiromitsu Tsuji, Youichi Tamai
The effect of replacement of negatively charged amino acid residues on the function of Na+ transport proteins of the salt-tolerant yeast Zygosaccharomyces rouxii was examined by heterologous expression of mutant proteins in a strain of Saccharomyces cerevisiae, RH16.6, that lacks native Na+-ATPase activity due to null mutations of ENA1, ENA2, ENA3, and ENA4. Mutants of Na+/H+ antiporter gene (ZrSOD2) and Na+-ATPase gene (ZrENA1) of Z. rouxii were generated by site-directed mutagenesis. The significance of two aspartic residues arranged in tandem (D265 and D266) was demonstrated in Z...
March 19, 2002: FEMS Microbiology Letters
F W Leak, S Tove, L W Parks
Wild-type yeast cells are unable to take up sterols from their growth media under aerobic conditions and are relatively resistant to monovalent cations. A yeast mutant (upc2-1) with a defect in the aerobic exclusion of sterols was found to have increased sensitivity to LiCl and NaCl. Although cation sensitivity has been reported for mutants that synthesize altered sterols, the mutant with upc2-1 continues to produce the normal sterol, ergosterol. The ENA2 gene was cloned on the basis of remediating the hypersensitivity to the monovalent cations...
February 1999: DNA and Cell Biology
K Blease, A Burke-Gaffney, P G Hellewell
1. Expression of cell adhesion molecules (CAM) on the lung microvascular endothelium is believed to play a key role in the recruitment of leukocytes in pulmonary inflammation. Moreover, regulation of CAM expression may be an important mechanism through which this inflammation may be controlled. Experimental evidence has suggested that combined phosphodiesterase (PDE) 3 and 4 inhibitors increase cyclic AMP levels within cells greater than inhibition of either isoenzyme alone. In the present study we assessed the effect of combinations of rolipram (PDE4 inhibitor), ORG 9935 (PDE3 inhibitor) and salbutamol (beta-agonist) on CAM expression and neutrophil or eosinophil adhesion to human lung microvascular endothelial cells (HLMVEC)...
May 1998: British Journal of Pharmacology
M A Bañuelos, A Rodríguez-Navarro
Two genes isolated from Schwanniomyces occidentalis, ENA1 and ENA2, encode P-type ATPases highly homologous to the Na-ATPases of Saccharomyces cerevisiae and complement the Na+ sensitivity of an S. cerevisiae mutant strain lacking its own Na-ATPases. The expression of both ENA1 and ENA2 was highly dependent on a high external pH, but whereas a high pH was sufficient for the expression of ENA2, the expression of ENA1 required a high pH and the presence of Na+. Disruption of ENA1 rendered the cells less tolerant to Na+ than the wild-type strain and decreased their capacity for Na+ extrusion...
January 16, 1998: Journal of Biological Chemistry
K M Schweitzer, A M Dräger, P van der Valk, S F Thijsen, A Zevenbergen, A P Theijsmeijer, C E van der Schoot, M M Langenhuijsen
The first step in the homing of hematopoietic progenitor cells (HPCs) from the peripheral blood to the bone marrow involves an adhesion molecule-dependent contact with human bone marrow endothelial cells (HBMECs). In the present study we describe the constitutive expression of two of these molecules, E-selectin and vascular cell adhesion molecule-1 (VCAM-1), on endothelial cells of hematopoietic tissues. Immunophenotypic analysis of tissue sections of hematopoietically active (human adult and fetal bone marrow, fetal spleen, fetal liver, and adult spleen with extramedullary hematopoiesis) and inactive tissues (human adult spleen, lymph node, appendix, and liver; and fetal lung and fetal intestine) revealed that E-selectin and VCAM-1 are selectively expressed on endothelial cells of adult and fetal hematopoietic organs...
January 1996: American Journal of Pathology
B Garciadeblas, F Rubio, F J Quintero, M A Bañuelos, R Haro, A Rodríguez-Navarro
The ENA2 gene encoding a P-type ATPase involved in Na+ and Li+ effluxes in Saccharomyces cerevisiae has been isolated. The putative protein encoded by ENA2 differs only in thirteen amino acids from the protein encoded by ENA1/PMR2. However, ENA2 has a very low level of expression and for this reason did not confer significant Li+ tolerance on a Li+ sensitive strain. ENA1 and ENA2 are the first two units of a tandem array of four highly homologous genes with probably homologous functions.
January 1993: Molecular & General Genetics: MGG
B C Hakkert, T W Kuijpers, J F Leeuwenberg, J A van Mourik, D Roos
Pretreatment of endothelial cells with cytokines enhances the adherence of leukocytes, a process that is mediated by surface proteins expressed on both cell types. A three-dimensional model system for the simultaneous determination of leukocyte adherence and migration was used to study the contribution of CD11/CD18, endothelial leukocyte-adhesion molecule-1 (ELAM-1) and VLA-4 in neutrophil and monocyte adherence to and migration through cytokine-activated endothelial cells. Pretreatment of endothelial cells for 4 hours with recombinant interleukin-1 beta (rIL-1 beta) was found to enhance neutrophil adherence and migration to a much greater extent than monocyte adherence and migration...
November 15, 1991: Blood
J F Leeuwenberg, T M Jeunhomme, W A Buurman
The role of ELAM-1 in the adhesion of monocytes to HUVEC, activated for 4h with TNF, was studied using MoAb ENA2 directed against ELAM-1. In a standard adhesion assay at 37 degrees C, F(ab')2 fragments of ENA2 did not, or weakly inhibited adhesion. When metabolic activity of the monocytes was reduced by (i) fixing the monocytes, (ii) performing the adhesion assay at 4 degrees C, and (iii) combining the forementioned conditions, the adhesion of the monocytes was strongly blocked by ENA2 and less effective or not by MoAb IB4 anti-CD18...
March 1992: Scandinavian Journal of Immunology
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