Publication: Implicación de los genes CDC 15 y NPKI en el fenotipo autolítico Lyti de Saccharomyces cerevisiae y aislamiento de dos genes heterólogos supresores
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2002
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Universidad Complutense de Madrid, Servicio de Publicaciones
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Abstract
La cepa mutante lyt1 de Saccharomyces cerevisiae presenta dos características fenotípicas independientes: un fenotipo cdc autolítico a 37 grados c. y un defecto en meiosis independiente de la temperatura. El fenotipo cdc es debido a una mutación en el orf del gen cdc15. Este hecho se confirmó por la clonación de dicho gen a partir de una genoteca genómica y se caracterizó como responsable de dicho fenotipo; el rescate del alelo cdc 15 de la cepa mutante lyt1; la transcripción de cdc15 se halla disminuida en la estirpe mutante. El defecto en esporulación se debe a una mutación en el marco abierto de lectura del gen npk1. Al igual que en el caso anterior, este hecho se confirmo por: el aislamiento y caracterización de npk1 a partir de una genoteca genómica; el rescate del alelo mutado de una estirpe lyt1, y por el estudio de su nivel de transcripción en la estirpe mutante. npr1p, de función desconocida hasta ahora, es imprescindible, en un fondo cdc15, para la consecución del ciclo meiótico en S. cerevisiae. Se han aislado, además, dos genes heterólogos (siti de S. pombe, y hlt1 humano) capaces de complementar el defecto en esporulación de una cepa lyt1
The Saccharomyces cerevisiae lyt1 mutant strain exhibits two independent phenotypic characteristics: an autolytic cdc phenotype at 37 degrees c. and a defect in meiosis independent of temperature. The cdc phenotype is due to a mutation in the orf of the cdc15 gene. This fact was confirmed by the cloning of said gene from a genomic library and was characterized as being responsible for said phenotype; rescue of the cdc 15 allele of the lyt1 mutant strain; cdc15 transcription is decreased in the mutant strain. The sporulation defect is due to a mutation in the open reading frame of the npk1 gene. As in the previous case, this fact was confirmed by: the isolation and characterization of npk1 from a genomic library; the rescue of the mutated allele of a lyt1 strain, and by studying its level of transcription in the mutant strain. npr1p, of unknown function until now, is essential, in a cdc15 background, for the achievement of the meiotic cycle in S. cerevisiae. In addition, two heterologous genes have been isolated (S. pombe siti and human hlt1) capable of complementing the defect in sporulation of a lyt1 strain
The Saccharomyces cerevisiae lyt1 mutant strain exhibits two independent phenotypic characteristics: an autolytic cdc phenotype at 37 degrees c. and a defect in meiosis independent of temperature. The cdc phenotype is due to a mutation in the orf of the cdc15 gene. This fact was confirmed by the cloning of said gene from a genomic library and was characterized as being responsible for said phenotype; rescue of the cdc 15 allele of the lyt1 mutant strain; cdc15 transcription is decreased in the mutant strain. The sporulation defect is due to a mutation in the open reading frame of the npk1 gene. As in the previous case, this fact was confirmed by: the isolation and characterization of npk1 from a genomic library; the rescue of the mutated allele of a lyt1 strain, and by studying its level of transcription in the mutant strain. npr1p, of unknown function until now, is essential, in a cdc15 background, for the achievement of the meiotic cycle in S. cerevisiae. In addition, two heterologous genes have been isolated (S. pombe siti and human hlt1) capable of complementing the defect in sporulation of a lyt1 strain
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Tesis de la Universidad Complutense de Madrid, Facultad de Farmacia, Departamento de Microbiología II, leída el 02-06-1995