Transcript analyses of cisplatin and Sky1 effects in Saccharomyces cerevisiae

ABSTRACT: Sky1 is a Saccharomyces cerevisiae rich serine-arginine (SR) protein-specific kinase and its enzymatic activity is essential in the cytotoxicity caused by cisplatin, although the molecular mechanisms supporting this function are not understood. We present a transcriptome analysis discriminating between RNA changes induced by cisplatin which are dependent or independent of the Sky1 function. The S. cerevisiae strain W303 (MATa ade2-1 can1-100 leu2-3,112 trp1-100 ura3- 52) and its derivative W303-M-NM-^Tsky1 previously described (RodrM-CM--guez- Lombardero et al., 2012) have been used in these analyses. Biological replicates of cultures and treatments were run in triplicate. The yeast cells were pre-cultured over night in 10mL of complete synthetic medium (SD) prepared as previously described (Zitomer and Hall, 1976). The following day the cells were inoculated at initial OD600 of 0.4 in 70 mL SD and grown in 250 mL Erlenmeyer flasks at 30 M-BM-:C and with agitation at 250 rpm. When cells reached OD600 of 0.6, the cultures from each strain were divided in two aliquots of 25 mL (control and cisplatin treatment). A stock solution of cisplatin 6 mM in dimethyl sulfoxide (DMSO) was prepared and the drug was added to the treated cultures at a final concentration of 600 microM. An equivalent volume of DMSO was added to the control cultures. The treatment was done at 30M-BM-:C and with agitation at 250 rpm during four hours in darkness. RNA was extracted from a number of cells corresponding to OD600 of 3 with the AurumTM Total RNA Mini Kit (Bio-Rad) and following the manufacturerM-bM-^@M-^Ys instructions. The RIN parameter (RNA Integrity Number) evaluated with the 2100 was near to the value 9 in all the samples. Three different RMA analyses were performed: Analysis 1: GSM1008483-GSM1008488 Analysis 2: GSM1008489-GSM1008494 Analysis 3: GSM1008495-GSM1008500

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: M Esperanza Cerdan 

PROVIDER: E-GEOD-41094 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress