RESUMO
Quorum sensing by farnesol in Candida albicans inhibits filamentation and may be directly related to its ability to cause both mucosal and systemic diseases. The Ras1-cyclic AMP signaling pathway is a target for farnesol inhibition. However, a clear understanding of the downstream effectors of the morphological farnesol response has yet to be unraveled. To address this issue, we screened a library for mutants that fail to respond to farnesol. Six mutants were identified, and the czf1Δ/czf1Δ mutant was selected for further characterization. Czf1 is a transcription factor that regulates filamentation in embedded agar and also white-to-opaque switching. We found that Czf1 is required for filament inhibition by farnesol under at least three distinct environmental conditions: on agar surfaces, in liquid medium, and when embedded in a semisolid agar matrix. Since Efg1 is a transcription factor of the Ras1-cyclic AMP signaling pathway that interacts with and regulates Czf1, an efg1Δ/efg1Δ czf1Δ/czf1Δ mutant was tested for filament inhibition by farnesol. It exhibited an opaque-cell-like temperature-dependent morphology, and it was killed by low farnesol levels that are sublethal to wild-type cells and both efg1Δ/efg1Δ and czf1Δ/czf1Δ single mutants. These results highlight a new role for Czf1 as a downstream effector of the morphological response to farnesol, and along with Efg1, Czf1 is involved in the control of farnesol-mediated cell death in C. albicans.
Assuntos
Candida albicans/metabolismo , Proteínas de Ligação a DNA/metabolismo , Farneseno Álcool/farmacologia , Proteínas Fúngicas/metabolismo , Percepção de Quorum , Fatores de Transcrição/metabolismo , Candida albicans/efeitos dos fármacos , Candida albicans/genética , Candida albicans/fisiologia , Proteínas de Ligação a DNA/genética , Proteínas Fúngicas/genética , Temperatura Alta , Mutação , Fatores de Transcrição/genéticaRESUMO
A scalable four-step synthesis of the ornithine transcarbamylase inhibitor N(5)-phosphonoacetyl-l-ornithine (PALO) is achieved through boroxazolidinone protection of ornithine. Investigations in the model organism Saccharomyces cerevisiae found that, in contrast to a previous report, PALO did not influence growth rate or expression of genes involved in arginine metabolism.
Assuntos
Arginina/metabolismo , Ornitina/análogos & derivados , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Regulação da Expressão Gênica , Ornitina/síntese química , Ornitina/química , Ornitina/farmacologia , Ornitina Carbamoiltransferase/antagonistas & inibidores , Ornitina Carbamoiltransferase/metabolismo , Saccharomyces cerevisiae/efeitos dos fármacos , Proteínas de Saccharomyces cerevisiae/genéticaRESUMO
Nonsense-mediated mRNA decay is a highly conserved pathway that degrades mRNAs with premature termination codons. These mRNAs include mRNAs transcribed from nonsense or frameshift alleles as well as wild-type mRNA with signals that direct ribosomes to terminate prematurely. This unit describes techniques to monitor steady-state mRNA levels, decay rates, and structural features of mRNAs targeted by this pathway, as well as in vivo analysis of nonsense suppression and allosuppression in the yeast Saccharomyces cerevisiae. Protocols for the structural features of mRNA include analysis of cap status, 5' and 3' untranslated region (UTR) lengths, and poly(A) tail length.
Assuntos
Degradação do RNAm Mediada por Códon sem Sentido , Saccharomyces cerevisiae/genética , Técnicas Genéticas , RNA Fúngico/químicaRESUMO
The objective of the research was to determine the optimum application rates of soybean peroxidase (SBP) plus peroxide (SBPP) for reducing odorous VOC emissions from swine manure. Industrial-grade SBP was applied in combination with liquid hydrogen peroxide (H(2)O(2)) or powdered calcium peroxide (CaO(2)) to standard phenolic solutions and swine manure, and emissions were measured in a wind tunnel. The primary odorant in the untreated manure was 4-methylphenol, which accounted for 68-81% of the odor activity value. At the optimum application rate of SBPP (50 g L(-1)), 4-methylphenol emissions were reduced from the swine manure by 62% (H(2)O(2)) and 98% (CaO(2)) after 24h (P<0.0001). The CaO(2) had a longer residence time, remaining effective for 48 h with 92% reduction in emission rates (P<0.0001), while H(2)O(2) was similar to the control at 48 h (P=0.28).