RESUMO
The URA3 gene of Debaryomyces hansenii, encoding orotidine 5'-phosphate decarboxylase enzyme, was isolated by complementation in the yeast Saccharomyces cerevisiae. The deduced amino acid sequence is highly similar to Ura3 proteins from other yeast and fungal species. Analysis of the region upstream of the coding sequence revealed the presence of AG-rich minisatellite DNA sequences. In addition, upstream of the DURA3 sequence, we have found the 3'-terminal of a gene encoding a GEA2-like protein.
Assuntos
Proteínas Fúngicas/genética , Repetições Minissatélites/genética , Proteínas de Saccharomyces cerevisiae , Saccharomycetales/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Genes Fúngicos , Teste de Complementação Genética , Fatores de Troca do Nucleotídeo Guanina/genética , Dados de Sequência Molecular , Saccharomycetales/fisiologia , Análise de Sequência de DNA , Cloreto de Sódio/farmacologiaRESUMO
The PBS2 gene encodes a MAP kinase kinase that plays a pivotal role in osmosensing signal-transduction pathway in the yeast Saccharomyces cerevisiae. Mutation in the PBS2 gene has a pleotropic effect. Besides being osmosensitive, pbs2 mutants show altered sensitivity to polymyxin B and calcofluor. Recent studies revealed that Pbs2p plays a different role in osmoadaptation and calcofluor sensitivity. We have isolated a gene homologous to PBS2 from the highly salt-tolerant yeast Debaryomyces hansenii by phenotypic complementation. DNA sequencing of the clone revealed that the gene encoded a protein of 683 amino acid residues. Like Pbs2p, this protein also has a proline-rich motif. Further characterization revealed that this gene could complement polymyxin B sensitivity but did not affect calcofluor sensitivity. Thus, it appeared that Pbs2p also has an independent role in these two physiological processes. The GenBank Accession No. of this sequence is AF371315.
Assuntos
Quinases de Proteína Quinase Ativadas por Mitógeno/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Mutação , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Saccharomycetales/genética , Sequência de Aminoácidos , Antibacterianos/farmacologia , Antifúngicos/farmacologia , Sequência de Bases , Benzenossulfonatos/farmacologia , Corantes Fluorescentes/farmacologia , Testes de Sensibilidade Microbiana , Quinases de Proteína Quinase Ativadas por Mitógeno/química , Dados de Sequência Molecular , Polimixina B/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/enzimologia , Saccharomycetales/enzimologia , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosRESUMO
The HOG1 gene encodes a MAP kinase that plays an essential role in maintaining water homeostasis in the yeast Saccharomyces cerevisiae. A gene homologous to S. cerevisiae HOG1 has been isolated from a highly salt-tolerant yeast, Debaryomyces hansenii, by phenotypic complementation. DNA sequencing of the clone revealed the presence of an open reading frame encoding a protein 387 amino acids long. The deduced amino acid sequence showed very high similarity with homologous genes identified from S. cerevisiae, Candida albicans and Zygosaccharomyces rouxii. In addition, it has also TGY motif characteristics of hyperosmolarity-activated MAP kinases. The Genbank Accession No. of this sequence is AF185278.