Transcriptomics,Genomics

Dataset Information

33

Transcriptomic analyses of IXR1 gene deletion in Saccharomyces cerevisiae and its increased resistance to cisplatin treatment.


ABSTRACT: Ixr1 is a transcriptional factor from Saccharomyces cerevisae with high affinity to cisplatin-DNA adducts through their two HMG-box DNA binding domains. Its transcriptional regulation 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 Ixr1 function. Overall design: The S. cerevisiae strain W303 (MATa ade2-1 can1-100 leu2-3,112 trp1-100 ura3- 52) and its derivative W303-Δsky1 previously described (Rodríguez- 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 ºC 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 30ºC 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 manufacturer’s instructions. The RIN parameter (RNA Integrity Number) evaluated with the 2100 was near to the value 9 in all the samples.

INSTRUMENT(S): [Yeast_2] Affymetrix Yeast Genome 2.0 Array

SUBMITTER: Angel Vizoso-Vázquez  

PROVIDER: GSE84569 | GEO | 2018-02-21

REPOSITORIES: GEO

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Publications

Ixr1 Regulates Ribosomal Gene Transcription and Yeast Response to Cisplatin.

Vizoso-Vázquez Ángel Á   Lamas-Maceiras Mónica M   González-Siso M Isabel MI   Cerdán M Esperanza ME  

Scientific reports 20180215 1


Ixr1 is a Saccharomyces cerevisiae HMGB protein that regulates the hypoxic regulon and also controls the expression of other genes involved in the oxidative stress response or re-adaptation of catabolic and anabolic fluxes when oxygen is limiting. Ixr1 also binds with high affinity to cisplatin-DNA adducts and modulates DNA repair. The influence of Ixr1 on transcription in the absence or presence of cisplatin has been analyzed in this work. Ixr1 regulates other transcriptional factors that respo  ...[more]

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