BioProject

Periodic fever syndrome is a disease that affects children mainly in the Mediterranean region, which has as one of the typical conditions of inflammation generalized by the body that can spontaneously regress. More...

Periodic fever syndrome is a disease that affects children mainly in the Mediterranean region, which has as one of the typical conditions of inflammation generalized by the body that can spontaneously regress. It has already been established that this pattern is presented by individuals carrying a mutant allele of the gene encoding the enzyme mevalonate kinase (MVK). This gene has an ortholog in the yeast Saccharomyces cerevisiae (ERG12) and mutations in this genes show phenotype that resemble those of the cells of these individuals. The present work aims to use this yeast as a biological model to analyze the role of the ERG12 gene (MVK) in the development of the disease. In this experiment, we used a wild type strain for the enzyme Erg12 (wt) and a genetically modified strain that decreases the activity of this enzyme (erg12-d) to 10%. The cells were cultured in synthetic medium containing glucose (reference medium) and each was re-inoculated and synthetic medium containing glucose or glycerol (erg12-d cells have growth deficiency in glycerol). By attaining D.O. value around 0.5 (600nm), the cells were collected for total RNA extraction and this was subjected to double-labeled microarray analysis in four combinations: erg12-d/wt and glycerol/glucose. Overall design: A MVK-positive parental strain and a MVK-deficient Damp mutant (showing 10% of parental enzyme activity) were cultivated in synthetic YNB medium containing glucose to mid-exponential growth phase, collected, washed and suspended in YNB containing glucose or glycerol as carbon source. After one hour of incubation, cells were collected for total RNA extraction. Following cRNA synthesis and labelling by two-color method, slides were hybridised and scanned. Two arrays, corresponding to biological duplicate, were used for each sample combination: parental glycerol (Cy5, test) x parental glucose (Cy3, reference); Damp glycerol (Cy5, test) x Damp glucose (Cy3 reference); Damp glucose (Cy5, test) x parental glucose (Cy3, reference); Damp glycerol (Cy5, test) x parental glycerol (Cy3, reference). Internal reference included RNA spike. Please note that there are two biological replicates for each sample comparison (ie.g. two raw data files per sample). However, the replicate data were combined and normalized together, resulting in 4 sample records. Less...