Skip to main content
Log in

Identification of new proteins related with cisplatin resistance in Saccharomyces cerevisiae

  • Biotechnologically relevant enzymes and proteins
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

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 modified protein abundance index (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 protein interactions. KEGG tool was used for metabolic pathway analysis. Data are available via ProteomeXchange with identifier PXD020665. The cisplatin-resistant strain showed 2.5 times more resistance than the wt strain for the inhibitory dose 50% (ID50) value (224 μg/ml vs 89.68 μg/ml) and 2.78 times more resistant for the inhibitory dose 90% (ID90) value (735.2 μg/ml vs 264.04 μg/ml). Multiple deregulated proteins were found in the glutathione and carbon metabolism, oxidative phosphorylation, proteasome, glycolysis and gluconeogenesis, glyoxylate metabolism, fatty acid degradation pathway, citric acid cycle, and ribosome. The most overexpressed proteins in the cisplatin-resistant strain were related to growth and metabolism (QCR2, QCR1, ALDH4, ATPB, ATPA, ATPG, and PCKA), cell structure (SCW10), and thermal shock (HSP26). The results suggest that these proteins could be involved in cisplatin resistance. The resistance acquisition process is complex and involves the activation of multiple mechanisms that interact together.

Key points

• Identification of new proteins/genes related to cisplatin resistance

• Increased expression of QCR2/QCR1/ALDH4/ATPB/ATPA/SCW10/HSP26/ATPG and PCKA proteins

• Multiple molecular mechanisms that interact together are involved in resistance

Graphical abstract

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (Perez-Riverol et al. 2019) partner repository with the dataset identifier PXD020665.

References

  • Aggarwal SK (1993) A histochemical approach to the mechanism of action of cisplatin and its analogues. J Histochem Cytochem 41:1053–1073

    Article  CAS  PubMed  Google Scholar 

  • Bodiga S, Vemuri PK, Bodiga VL (2018) Low Ctr1p, due to lack of Sco1p results in lowered cisplatin uptake and mediates insensitivity of rho0 yeast to cisplatin. J Inorg Biochem 187:14–24

    Article  CAS  PubMed  Google Scholar 

  • Branco P, Francisco D, Chambon C, Hébraud M, Arneborg N, Almeida MG, Caldeira J, Albergaria H (2014) Identification of novel GAPDH-derived antimicrobial peptides secreted by Saccharomyces cerevisiae and involved in wine microbial interactions. Appl Microbiol Biotechnol 98:843–853

    Article  CAS  PubMed  Google Scholar 

  • Brozovic A, Ambriović-Ristov A, Osmak M (2010) The relationship between cisplatin-induced reactive oxygen species, glutathione, and BCL-2 and resistance to cisplatin. Crit Rev Toxicol 40:347–359

    Article  CAS  PubMed  Google Scholar 

  • Casares C, Ramírez-Camacho R, Trinidad A, Roldán A, Jorge E, García-Berrocal JR (2012) Reactive oxygen species in apoptosis induced by cisplatin: review of physiopathological mechanisms in animal models. Eur Arch Otorhinolaryngol 269:2455–2459

    Article  PubMed  Google Scholar 

  • Chao CC (1996) Molecular basis of cis-diamminedichloroplatinum (II) resistance: a review. J Formos Med Assoc 95:893–900

    CAS  PubMed  Google Scholar 

  • Chen RE, Thorner J (2007) Function and regulation in MAPK signaling pathways: lessons learned from the yeast Saccharomyces cerevisiae. Biochim Biophys Acta 1773:1311–1340

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Costa AR, Machado N, Rego A, Sousa MJ, Côrte-Real M (2019) Proteasome inhibition prevents cell death induced by the chemotherapeutic agent cisplatin downstream of DNA damage. DNA Repair 73:28–33

    Article  CAS  PubMed  Google Scholar 

  • Dasari S, Tchounwou PB (2014) Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol 740:364–378

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dempke W, Voigt W, Grothey A, Hill BT, Schmoll HJ (2000) Cisplatin resistance and oncogenes-a review. Anti-Cancer Drugs 11:225–236

    Article  CAS  PubMed  Google Scholar 

  • Desoize B (2002) Cancer and metals and metal compounds. Critical Rev Oncol/Hematol 42:1–3

    Article  Google Scholar 

  • Galluzzi L, Vitale I, Senovilla L, Eisenberg T, Carmona-Gutierrez D, Vacchelli E, Robert T, Ripoche H, Jägemann N, Paccard C, Servant N, Hupé P, Lazar V, Dessen P, Barillot E, Zischka H, Madeo F, Kroemer G (2012) Independent transcriptional reprogramming and apoptosis induction by cisplatin. Cell Cycle 11:3472–3480

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gibson N, McAlister-Henn L (2003) Physical and genetic interactions of cytosolic malate dehydrogenase with other gluconeogenic enzymes. J Biol Chem 278:25628–25636

    Article  CAS  PubMed  Google Scholar 

  • Huang RY, Eddy M, Vujcic M, Kowalski D (2005) Genome-wide screen identifies genes whose inactivation confer resistance to cisplatin in Saccharomyces cerevisiae. Cancer Res 65:5890–5897

    Article  CAS  PubMed  Google Scholar 

  • Ishihama Y, Oda Y, Tabata T, Sato T, Nagasu T, Rappsilber J, Mann M (2005) Exponentially modified protein abundance index (emPAI) for estimation of absolute protein amount in proteomics by the number of sequenced peptides per protein. Mol Cell Proteomics 4:1265–1272

    Article  CAS  PubMed  Google Scholar 

  • Izaguirre DI, Ng CW, Kwan SY, Kun EH, Tsang YTM, Gershenson DM, Wong KK (2020) The role of GDF15 in regulating the canonical pathways of the tumor microenvironment in wild type p53 ovarian tumor and its response to chemotherapy. Cancers 12(10):E3043

    Article  PubMed  Google Scholar 

  • Kanehisa M, Goto S (2000) KEGG: Kyoto Encyclopedia of Genes and Genomes. Nucleic Acids Res 28:27–30 [pubmed] [doi]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kanehisa M, Sato Y (2020) KEGG Mapper for inferring cellular functions from protein sequences. Protein Sci 29:28–35

    Article  CAS  PubMed  Google Scholar 

  • Klein TE, Chang JT, Cho MK, Easton KL, Fergerson R, Hewett M, Lin Z, Liu Y, Liu S, Oliver DE, Rubin DL, Shafa F, Stuart JM, Altman RB (2001) Integrating genotype and phenotype information: an overview of the PharmGKB project. Pharmacogenetics Research Network and Knowledge Base. Pharmacogenomics J 1:167–170

    Article  CAS  PubMed  Google Scholar 

  • Kowalski D, Pendyala L, Daignan-Fornier B, Howell SB, Huang RY (2008) Dysregulation of purine nucleotide biosynthesis pathways modulates cisplatin cytotoxicity in Saccharomyces cerevisiae. Mol Pharmacol 74:1092–1100

    Article  CAS  PubMed  Google Scholar 

  • Larriba G, Basco RD, Andaluz E, Luna-Arias JP (1993) Yeast exoglucanases. Where redundancy implies necessity. Arch Med Res 24:293–299

    CAS  PubMed  Google Scholar 

  • Liao C, Hu B, Arno MJ, Panaretou B (2007) Genomic screening in vivo reveals the role played by vacuolar H+ ATPase and cytosolic acidification in sensitivity to DNA-damaging agents such as cisplatin. Mol Pharmacol 71:416–425

    Article  CAS  PubMed  Google Scholar 

  • Mariani D, Castro FA, Almeida LG, Fonseca FL, Pereira MD (2014) Protection against cisplatin in calorie-restricted Saccharomyces cerevisiae is mediated by the nutrient-sensor proteins Ras2, Tor1, or Sch9 through its target glutathione. FEMS Yeast Res 14:1147–1159

    Article  CAS  PubMed  Google Scholar 

  • Mercado-Sáenz S, López-Díaz B, Sendra-Portero F, Martínez-Morillo M, Ruiz-Gómez MJ (2017) Inactivation of RAD52 and HDF1 DNA repair genes leads to premature chronological aging and cellular instability. J Biosci 42:219–230

    Article  PubMed  Google Scholar 

  • Montazeri V, Ghahremani MH, Montazeri H, Hasanzad M, Safavi M, Ayati M, Chehrazi M, Moghaddam BA, Ostad SN (2020) A preliminary study of NER and MMR pathways involved in chemotherapy response in bladder transitional cell carcinoma: Impact on progression free survival. Iranian J Pharm Res 19(1):355–365

    CAS  Google Scholar 

  • Ozben T (2007) Oxidative stress and apoptosis: impact on cancer therapy. J Pharm Sci 96:2181–2196

    Article  CAS  PubMed  Google Scholar 

  • Perez-Riverol Y, Csordas A, Bai J, Bernal-Llinares M, Hewapathirana S, Kundu DJ, Inuganti A, Griss J, Mayer G, Eisenacher M, Pérez E, Uszkoreit J, Pfeuffer J, Sachsenberg T, Yilmaz S, Tiwary S, Cox J, Audain E, Walzer M, Jarnuczak AF, Ternent T, Brazma A, Vizcaíno JA (2019) The PRIDE database and related tools and resources in 2019: improving support for quantification data. Nucleic Acids Res 47(D1):D442–D450

    Article  CAS  PubMed  Google Scholar 

  • Qiao XL, Zhong ZL, Dong Y, Gao F (2020) LncRNA HMGA1P4 promotes cisplatin-resistance in gastric cancer. Eur Rev Med Pharmacol Sci 24(17):8830–8836

    PubMed  Google Scholar 

  • Rak M, Tzagoloff A (2009) F1-dependent translation of mitochondrially encoded Atp6p and Atp8p subunits of yeast ATP synthase. Proc Natl Acad Sci U S A 106:18509–18514

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Reinders J, Wagner K, Zahedi RP, Stojanovski D, Eyrich B, Van der Laan M, Rehling P, Sickmann A, Pfanner N, Meisinger C (2007) Profiling phosphoproteins of yeast mitochondria reveals a role of phosphorylation in assembly of the ATP synthase. Mol Cell Proteomics 6:1896–1906

    Article  CAS  PubMed  Google Scholar 

  • Ruiz-Gómez MJ, Martínez-Morillo M (2006) Iron (III) chloride hexahydrate does not enhance methotrexate cytotoxicity on Saccharomyces cerevisiae. Chemotherapy 52:226–230

    Article  PubMed  Google Scholar 

  • Ruiz-Gómez MJ, Merino-Moyano MD, Cebrián-Martín MG, Prieto-Barcia MI, Martínez-Morillo M (2008) No effect of 50 Hz 2.45 mT magnetic field on the potency of cisplatin, mitomycin C and methotrexate in S. cerevisiae. Electromagn Biol Med 27:289–297

    Article  PubMed  Google Scholar 

  • Saad SY, Najjar TA, Alashari M (2004) Role of non-selective adenosine receptor blockade and phosphodiesterase inhibition in cisplatin-induced nephrogonadal toxicity in rats. Clin Exp Pharmacol Physiol 31:862–867

    Article  CAS  PubMed  Google Scholar 

  • Schenk PW, Brok M, Boersma AW, Brandsma JA, Den Dulk H, Burger H, Stoter G, Brouwer J, Nooter K (2003) Anticancer drug resistance induced by disruption of the Saccharomyces cerevisiae NPR2 gene: a novel component involved in cisplatin-and doxorubicin-provoked cell kill. Mol Pharmacol 64:259–268

    Article  CAS  PubMed  Google Scholar 

  • Schoch S, Gajewski S, Rothfuβ J, Hartwig A, Köberle B (2020) Comparative study of the mode of action of clinically approved platinum-based chemotherapeutics. Int J Mol Sci 21:6928

    Article  CAS  PubMed Central  Google Scholar 

  • Siddik ZH (2003) Cisplatin: mode of cytotoxic action and molecular basis of resistance. Oncogene 22:7265–7279

    Article  CAS  PubMed  Google Scholar 

  • Stewart DJ (2007) Mechanisms of resistance to cisplatin and carboplatin. Crit Rev Oncol Hematol 63:12–31

    Article  PubMed  Google Scholar 

  • Szklarczyk D, Morris JH, Cook H, Kuhn M, Wyder S, Simonovic M, Santos A, Doncheva NT, Roth A, Bork P, Jensen LJ, von Mering C (2017) The STRING database in 2017: quality-controlled protein-protein association networks, made broadly accessible. Nucleic Acids Res 45:D362–D3d8

    Article  CAS  PubMed  Google Scholar 

  • Tkach JM, Yimit A, Lee AY, Riffle M, Costanzo M, Jaschob D, Hendry JA, Ou J, Moffat J, Boone C, Davis TN, Nislow C, Brown GW (2012) Dissecting DNA damage response pathways by analysing protein localization and abundance changes during DNA replication stress. Nat Cell Biol 14:966–976

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tsvetanova NG, Klass DM, Salzman J, Brown PO (2010) Proteome-wide search reveals unexpected RNA-binding proteins in Saccharomyces cerevisiae. PLoS One 5:e12671

    Article  PubMed  PubMed Central  Google Scholar 

  • Tzagoloff A, Wu MA, Crivellone M (1986) Assembly of the mitochondrial membrane system. Characterization of COR1, the structural gene for the 44-kilodalton core protein of yeast coenzyme QH2-cytochrome c reductase. J Biol Chem 261:17163–17169

    Article  CAS  PubMed  Google Scholar 

  • White HE, Orlova EV, Chen S, Wang L, Ignatiou A, Gowen B, Stromer T, Franzmann TM, Haslbeck M, Buchner J, Saibil HR (2006) Multiple distinct assemblies reveal conformational flexibility in the small heat shock protein Hsp26. Structure 14:1197–1204

    Article  CAS  PubMed  Google Scholar 

  • Wu ZZ, Lu HP, Chao CCK (2010) Identification and functional analysis of genes which confer resistance to cisplatin in tumor cells. Biochem Pharmacol 80:262–276

    Article  CAS  PubMed  Google Scholar 

  • Yin Z, Hatton L, Brown AJ (2000) Differential post-transcriptional regulation of yeast mRNAs in response to high and low glucose concentrations. Mol Microbiol 35:553–565

    Article  CAS  PubMed  Google Scholar 

  • Yuan L, Yu WM, Qu CK (2003) DNA damage-induced G2/M checkpoint in SV40 large T antigen-immortalized embryonic fibroblast cells requires SHP-2 tyrosine phosphatase. J Biol Chem 278:42812–42820

    Article  CAS  PubMed  Google Scholar 

  • Zeitlinger J, Simon I, Harbison CT, Hannett NM, Volkert TL, Fink GR, Young RA (2003) Program-specific distribution of a transcription factor dependent on partner transcription factor and MAPK signaling. Cell 113:395–404

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We express our gratitude to Dr Anna A. Friedl (Department of Radiation Oncology, Ludwig-Maximilians-Universität München, Munich, Germany) for kindly providing the yeast strain. We thank Ms. L. Gil Carmona (University of Málaga, Spain) for her technical assistance in yeast culture. Thanks to Kanehisa Laboratories for permission to reproduce the images generated with KEGG pathway enrichment analysis tool.

Funding

This work was supported by the “Plan Andaluz de Investigación, Desarrollo e Innovación (PAIDI); Junta de Andalucía”, code CTS-181.

Author information

Authors and Affiliations

Authors

Contributions

MJRG conceived, designed, and supervised the study. AMBM, SMS, CC, BLD, and FSP conducted the experiments. AMBM, SMS, CC, BLD, FSP, and MJRG analyzed the data. AMBM, CC, and MJRG designed and generated the figures. AMBM, CC, and MJRG wrote the manuscript. All authors read and approved the manuscript.

Corresponding author

Correspondence to Miguel J. Ruiz-Gómez.

Ethics declarations

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Consent to participate

Not applicable.

Consent for publication

All listed authors have approved the manuscript before submission, including the names and order of authors.

Conflict of interest

The authors declare no competing interests.

Permission

Permission was obtained from Kanehisa Laboratories to reproduce the images generated with KEGG pathway enrichment analysis tool.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

ESM 1

(PDF 2455 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Burgos-Molina, A.M., Mercado-Sáenz, S., Cárdenas, C. et al. Identification of new proteins related with cisplatin resistance in Saccharomyces cerevisiae. Appl Microbiol Biotechnol 105, 1965–1977 (2021). https://doi.org/10.1007/s00253-021-11137-w

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-021-11137-w

Keywords

Navigation