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Over-expression of CTR1 delta-300 alters element and transcription profiles in yeast

ABSTRACT: In an approach to generate Saccharomyces cerevisiae strains with increased intracellular copper amounts for technical applications we over-expressed the copper transporter CTR1 and a variant of CTR1 with a truncation in the C-terminus after the 300 amino acids (CTR1 delta-300). We determined the copper sensitivity of the generated strains and used inductively coupled plasma spectrometry (ICP-OES and ICP-MS) analysis to investigate the effects of over-expression of both constructs under excess copper on the cellular content of different elements in S. cerevisiae. In addition, we ran DNA microarray analysis to obtain the gene expression profile under the changed element contents. Over-expression of CTR1 increased the copper content in the cells up to 160% and 78 genes were differentially regulated. Over-expression of the truncated CTR1 delta-300 resulted in an increased copper, iron and zinc content of more than 200% and 980 genes showed differential expression. We found that transition metal ion homeostasis was disrupted in CTR1 delta-300 over-expressing strains under excess copper and that this was combined with a transcriptional remodelling of cellular processes Transcription profiling was performed in Saccharomyces cerevisiae BY4741 (referred to as wild type) over-expressing CTR1 and in the ctr1- deletion strain BY4741 delta-ctr1 overexpressing CTR1 delta-300 after 7 hours of culture under excess copper (0.02 mM CuCl2). Control strains contained the corresponding empty vector. Three independent experiments were performed and 12 samples were analyzed.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Astrid Schuller

PROVIDER: E-GEOD-44043 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Over-expression of CTR1 delta-300 alters element and transcription profiles in yeast

Project description:In an approach to generate Saccharomyces cerevisiae strains with increased intracellular copper amounts for technical applications we over-expressed the copper transporter CTR1 and a variant of CTR1 with a truncation in the C-terminus after the 300 amino acids (CTR1 delta-300). We determined the copper sensitivity of the generated strains and used inductively coupled plasma spectrometry (ICP-OES and ICP-MS) analysis to investigate the effects of over-expression of both constructs under excess copper on the cellular content of different elements in S. cerevisiae. In addition, we ran DNA microarray analysis to obtain the gene expression profile under the changed element contents. Over-expression of CTR1 increased the copper content in the cells up to 160% and 78 genes were differentially regulated. Over-expression of the truncated CTR1 delta-300 resulted in an increased copper, iron and zinc content of more than 200% and 980 genes showed differential expression. We found that transition metal ion homeostasis was disrupted in CTR1 delta-300 over-expressing strains under excess copper and that this was combined with a transcriptional remodelling of cellular processes

2014-01-28 | GSE44043 | GEO

Saccharomyces cerevisiae

Project description:Over-expression of CTR1 delta-300 alters element and transcription profiles in yeast

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