RAD51 OVEREXPRESSION AND RESISTANCE TO GENOTOXIC AGENTS. A STUDY IN THE FISSION YEAST SCHIZOSACCHAROMYCES POMBE

Authors

  • Raluca Stefania STANESCU University of Medicine and Pharmacy “Grigore T. Popa” –Iasi
  • Gabriela BORDEIANU University of Medicine and Pharmacy “Grigore T. Popa” –Iasi
  • B. STOICA University of Medicine and Pharmacy “Grigore T. Popa” –Iasi
  • Didona UNGUREANU University of Medicine and Pharmacy “Grigore T. Popa” –Iasi
  • M. RUSU University of Medicine and Pharmacy “Grigore T. Popa” –Iasi
  • Elena Petrescu- DANILA University of Medicine and Pharmacy “Grigore T. Popa” –Iasi

Abstract

Background: Many cancer cell lines have been found to overexpress the recombinase Rad51. The overexpression is associated with increased invasive potential and resistance to DNA-damaging therapeutic agents. This has been attributed to an increased capacity of cells overexpressing Rad51 to repair DNA lesions or to a genetic stabilization of the genome. Aim: As the explanations are somewhat controversial, we attempted to reproduce overexpression in the unicellular eukaryote Schizosaccharomyces pombe to have a simpler tool to study the problem of Rad51 overexpression and its induced resistance to DNA-damaging agents. Methods: We used the nmt1 promoter inserted upstream of rad51gene to induce its overexpression and studied the phenotype of the transformed strain, especially its sensitivity to camptothecin and hydroxyurea.  Results: We found that overexpression induced sensitivity to the two drugs even when it was associated with the deletion of a recombination mediator rad22/rad52 gene.  However, when overexpression was associated with the deletion of the helicase-encoding fbh1 gene, the sensitivity to camptothecin was diminished.

Author Biographies

  • Raluca Stefania STANESCU, University of Medicine and Pharmacy “Grigore T. Popa” –Iasi

    Faculty of Medicine
    Department of Biochemistry

  • Gabriela BORDEIANU, University of Medicine and Pharmacy “Grigore T. Popa” –Iasi

    Faculty of Medicine
    Department of Biochemistry

  • B. STOICA, University of Medicine and Pharmacy “Grigore T. Popa” –Iasi

    Faculty of Medicine
    Department of Biochemistry

  • Didona UNGUREANU, University of Medicine and Pharmacy “Grigore T. Popa” –Iasi

    Faculty of Medicine
    Department of Biochemistry

  • M. RUSU, University of Medicine and Pharmacy “Grigore T. Popa” –Iasi

    Faculty of Medicine
    Department of Biochemistry

  • Elena Petrescu- DANILA, University of Medicine and Pharmacy “Grigore T. Popa” –Iasi

    Faculty of Medicine
    Department of Biochemistry

References

1. Krejci L, Altmannova V, Spirek M, Zhao X. Homologous recombination and its regulation. Nucleic Acids Res 2012; 40: 5795-5818.
2. Tsuzuki T, Fujii Y, Sakumi K et al. Targeted disruption of the Rad51 gene leads to lethality in em-bryonic mice. Proc Natl Acad Sci U S A 1996; 93: 6236–6240.
3. Klein HL. The consequences of Rad51 overexpression for normal and tumor cells. DNA Repair (Amst) 2008; 7(5): 686–693.
4. Raderschall E, Stout K, Freier S et al. Elevated levels of Rad51 recombination protein in tumor cells. Cancer Res 2002; 62: 219–225.
5. Hine CM, Seluanov A, Gorbunova V. Use of the Rad51 promoter for targeted anti-cancer therapy. Proc Natl Acad Sci USA 2008; 105: 20810-20815.
6. Hannay JA, Liu J, Zhu QS et al. Rad51 overexpression contributes to chemoresistance in human soft tissue sarcoma cells: a role for p53/activator protein 2 transcriptional regulation. Mol Cancer Ther 2007; 6: 1650–1660.
7. Slupianek A, Schmutte C, Tombline G et al. BCR/ABL regulates mammalian RecA homologs, resulting in drug resistance. Mol Cell 2001; 8: 795–806.
8. Powell SN, Kachnic LA. Roles of BRCA1 and BRCA2 in homologous recombination, DNA replica-tion fidelity and the cellular response to ionizing radiation. Oncogene 2003; 22, 5784–5791
9. Schild D, Wiese C. Overexpression of Rad51 suppresses recombination defects: a possible mechanism to reverse genomic instability. Nucl Acid Res 2010; 38: 1061-1070.
10. Brown ET, Holt J. Rad51 overexpression rescues radiation resistance in BRCA2-defective cancer cells. Mol Carcinog 2009; 48: 105–109.
11. Yang Z, Waldman AS, Wyatt MD. Expression and regulation of RAD51 mediate cellular responses to chemotherapeutics. Biochem Pharmacol 2012; 83: 741-746.
12. Kim WJ, Lee H, Park EJ, Park JK, Park SD. Gain- and loss-of-function of Rhp51, a Rad51 homolog in fission yeast, reveals dissimilarities in chromosome integrity. Nucleic Acids Res 2001; 29: 1724–1732.
13. Osman F, Dixon J, Barr AR, Whitby MC. The F-Box DNA Helicase Fbh1 Prevents Rhp51-Dependent Recombination without Mediator Proteins. Moll Cell Biol 2005; 25(18): 8084-8096.
14. Werler PJH, Hartsuiker E, Carr AM. A simple Cre-lox metod for chromosomal N-terminal tagging of essential and non-essential Schizosaccharomyces pombe genes. Gene 2003; 304: 133-141.
15. Petrescu-Danila E, Voicu PM, Poitelea M, Stanescu R, Stoica B, Rusu M. Method for protein tagging in the fission yeast Schizosaccharomyces pombe. Rev Med Chir Soc Med Nat Iasi 2006; 110: 403-408.
16. Bahler J, Wu JQ, Longtine MS et al. Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Yeast 1998; 14: 943-951.
17. Feng Z, Scott SP, Bussen W. et al., Rad52 inactivation is synthetically lethal with BRCA2 deficiency. Proc Natl Acad Sci U S A 2011; 108(2): 686-691.
18. Lorentz A, Osman F, Folkyte V, Sofueva S, Whitby MC. Fbh1 limits Rad51-dependent recombination at blocked replication forks. Mol and Cell Biol 2009; 29: 4742-4756.
19. Fugger K, Mistrik M, Danielsen JR et al. Human Fbh1 helicase contributes to genome maintenance via pro- and anti- recombinase activities. J. Cell Biol 2009; 186: 655-663.

Additional Files

Published

2018-05-15