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1.
Mol Biol Cell ; 25(1): 196-211, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24196832

RESUMO

The Saccharomyces cerevisiae protein kinase Sch9 is an in vitro and in vivo effector of sphingolipid signaling. This study examines the link between Sch9 and sphingolipid metabolism in S. cerevisiae in vivo based on the observation that the sch9Δ mutant displays altered sensitivity to different inhibitors of sphingolipid metabolism, namely myriocin and aureobasidin A. Sphingolipid profiling indicates that sch9Δ cells have increased levels of long-chain bases and long-chain base-1 phosphates, decreased levels of several species of (phyto)ceramides, and altered ratios of complex sphingolipids. We show that the target of rapamycin complex 1-Sch9 signaling pathway functions to repress the expression of the ceramidase genes YDC1 and YPC1, thereby revealing, for the first time in yeast, a nutrient-dependent transcriptional mechanism involved in the regulation of sphingolipid metabolism. In addition, we establish that Sch9 affects the activity of the inositol phosphosphingolipid phospholipase C, Isc1, which is required for ceramide production by hydrolysis of complex sphingolipids. Given that sphingolipid metabolites play a crucial role in the regulation of stress tolerance and longevity of yeast cells, our data provide a model in which Sch9 regulates the latter phenotypes by acting not only as an effector but also as a regulator of sphingolipid metabolism.


Assuntos
Ceramidas/biossíntese , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/enzimologia , Antifúngicos/farmacologia , Depsipeptídeos/farmacologia , Farmacorresistência Fúngica , Ácidos Graxos Monoinsaturados/farmacologia , Regulação Fúngica da Expressão Gênica , Técnicas de Inativação de Genes , Testes de Sensibilidade Microbiana , Viabilidade Microbiana , Proteínas Serina-Treonina Quinases/genética , Transporte Proteico , Saccharomyces cerevisiae/efeitos dos fármacos , Proteínas de Saccharomyces cerevisiae/genética , Esfingolipídeos/metabolismo , Esfingosina/análogos & derivados , Esfingosina/farmacologia , Transcrição Gênica , Fosfolipases Tipo C/metabolismo
2.
Anal Bioanal Chem ; 390(2): 783-93, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17985118

RESUMO

Burned greasy deposits were found inside shells of the large Nile bivalve Chambardia rubens, excavated in an eight- to tenth- century AD church of the Coptic monastery of Bawit, Egypt, and supposedly used as oil lamps. The residues were subjected to a combination of chromatographic residue analysis techniques. The rather high concentrations of unsaturated fatty acids, as analysed by gas chromatography (GC) in the methylated extract, suggest the presence of a vegetal oil. Analysis of the stable carbon isotopes (delta 13C values) of the methyl esters also favoured plants over animals as the lipid source. In the search for biomarkers by GC coupled to mass spectrometry on a silylated extract, a range of diacids together with high concentrations of 13,14-dihydroxydocosanoate and 11,12-dihydroxyeicosanoate were found. These compounds are oxidation products of erucic acid and gondoic acid, which are abundantly present in seeds of Brassicaceae plants. Liquid chromatography coupled to mass spectrometry analysis showed low concentrations of unaltered triglycerides, but revealed sizeable amounts of triglycerides with at least one dihydroxylated acyl chain. The unusual preservation of dihydroxylated triglycerides and alpha,omega-dicarboxylic acids can be related to the dry preservation conditions. Analysis of the stereoisomers of the dihydroxylated fatty acids allows one to determine whether oxidation took place during burning of the fuel or afterwards. The results prove that the oil of rapeseed (Brassica napus L.) or radish (Raphanus sativus L.) was used as illuminant in early Islamic Egypt, and that not only ceramic lamps but also mollusk shells were used as fuel containers.


Assuntos
Bivalves , Brassicaceae/química , Cristianismo , Óleos de Plantas/análise , Sementes/química , Animais , Egito , Iluminação , Óleos de Plantas/química , Rios , Fatores de Tempo
3.
FEBS Lett ; 580(7): 1903-7, 2006 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-16527275

RESUMO

The antifungal plant defensin DmAMP1 interacts with fungal sphingolipids of mannosyldiinositolphosphorylceramide (M(IP)2C) class. We screened a Saccharomyces cerevisiae transposon (Tn) mutant library against DmAMP1 and identified one DmAMP1-resistant mutant with the Tn inserted in the M(IP)2C biosynthesis gene IPT1 (DmTn11) and one DmAMP1-hypersensitive mutant with the Tn inserted in rDNA (HsTnII). However, tetrad analysis pointed to HsTnII as a spontaneous mutant. Apparently, membranes of DmTn11 lack M(IP)2C, whereas membranes of HsTnII have increased M(IP)2C levels. In addition, DmTn11 and HsTnII are characterized by increased and reduced oxidative stress resistance/chronological life-span (CL), respectively. A putative involvement of M(IP)2C in oxidative stress and CL in yeast is discussed.


Assuntos
Defensinas/fisiologia , Glicoesfingolipídeos/fisiologia , Estresse Oxidativo , Oxigênio/metabolismo , Leveduras/citologia , Glicoesfingolipídeos/análise , Mutação , Plantas/imunologia , Saccharomyces cerevisiae/citologia , Saccharomyces cerevisiae/genética
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