Dataset Information

Identification of new proteins related with cisplatin resistance in S. cerevisiae

ABSTRACT: The aim of this study is to select a cisplatin resistant Saccharomyces cerevisiae strain as a model organism to look for new molecular markers of cisplatin resistance and the identification of mechanisms/interactions involved. A cisplatin resistant S. cerevisiae strain was obtained after continuous exposure to the drug during 80 days. Then, total protein extraction, purification and identification were carried out, in wild type (wt) and resistant strains, by tandem mass spectrometry using a "nano HPLC-ESI-MS/MS" ion trap system. The increase in emPAI (resistant vs wt strains) was calculated to study the increase in protein expression. "Genemania" software (http://www.Genemania.org/) was used to compare the effects, functions and interactions of proteins. The selected cisplatin resistant strain showed 2.5 times more resistance than the wt strain (for the ID50 value) and 2.78 times more resistant for the ID90 value. The long-term exposure to cisplatin induced resistance in S. cerevisiae, obtaining an increased expression of QCR2, QCR1, ALDH4, ATPB, ATPA, SCW10, HSP26, ATPG, and PCKA proteins. The overexpression of the above-mentioned proteins suggests that they could be involved in cisplatin resistance. The resistance acquisition process is complex and involves the activation of multiple mechanisms that interact together.

INSTRUMENT(S): DB source UniProt, Mascot, Bruker Daltonics amaZon series, ProteinScape, emPAI value, DB source NCBI

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Miguel Ruiz-Gomez

LAB HEAD: Miguel J. Ruiz-Gómez

PROVIDER: PXD020665 | Pride | 2021-02-25

REPOSITORIES: Pride

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Publications

Identification of new proteins related with cisplatin resistance in Saccharomyces cerevisiae.

Burgos-Molina Antonio M AM Mercado-Sáenz Silvia S Cárdenas Casimiro C López-Díaz Beatriz B Sendra-Portero Francisco F Ruiz-Gómez Miguel J MJ

Applied microbiology and biotechnology 20210212 5

The aim of this study is to select a cisplatin-resistant Saccharomyces cerevisiae strain to look for new molecular markers of resistance and the identification of mechanisms/interactions involved. A resistant strain was obtained after 80 days of cisplatin exposure. Then, total protein extraction, purification, and identification were carried out, in wild-type (wt) and resistant strains, by tandem mass spectrometry using a "nano HPLC-ESI-MS/MS" ion trap system. The increase in the exponentially m ...[more]

PMID: 33576883

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Project description:Understanding the mechanism of resistance in platinum-based regimens for the treatment of high-grade serous ovarian cancer (HGSOC) is important for identifying new therapeutic targets to improve the clinical outcome of ovarian cancer patients. Mass spectrometry-based proteomic strategy was applied to spheroidal cisplatin sensitive and resistant HGSOC generated cell lines in the absence and presence of cisplatin drug. A complete expressed HGSOC proteome and phosphoproteome was characterized in cisplatin sensitive and resistant HGSOC cell lines providing insight into the mechanism of resistance development. PCA analysis showed that phosphorylation of a few proteins provides better classification than the whole proteome of the cellular subtypes. Specifically, a distinctive phosphoproteomic signature between cisplatin sensitive and resistant cell lines in the absence of drug was observed. This same phosphoproteomic signature was observed in our cisplatin sensitive cell line in the absence and presence of drug, indicating a vital role for phosphorylation of proteins in resistance development to cisplatin. The most phosphorylated protein was sequestosome (p62/SQSTM1). Differential expressions of apoptosis by the prognostic factor ratio of Bcl-2/Bax and autophagy, known to be regulated by p62/SQSTM1, was validated in the proteome data and by western blot analysis. A significant increase in apoptosis in the presence of cisplatin was observed in only the sensitive cell line while autophagy revealed increased expression in the resistant relative to sensitive cell line. Furthermore, site specific phosphorylation on 20 modified residues of sequestosome was characterized. Elevated expression of phosphorylation of sequestosome in resistant HGSOC cell lines was validated with western blot analysis. Here, we propose phosphorylation of sequestosome to be a marker and key in cisplatin resistance development in HGOSC ovarian cancers by shuttling ubiquitinated proteins to the autophagy pathway and influencing down-regulation of apoptosis.

Dataset Information

Identification of new proteins related with cisplatin resistance in S. cerevisiae

Publications

Identification of new proteins related with cisplatin resistance in Saccharomyces cerevisiae.

Similar Datasets

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