Catalase T-Deficient Fission Yeast Meiocytes Show Resistance to Ionizing Radiation
Abstract
:1. Introduction
2. Materials and Methods
2.1. Yeast Strains and Their Construction
2.2. Cell Culture for Meiotic Time-Courses
2.3. Spot Assays
2.4. X Irradiation
2.5. H2O2 Treatment
2.6. Detection of Reactive Oxygen Species
2.7. Live-Cell Imaging and Image Analysis
2.8. Statistics
3. Results
3.1. Mutants of ctt1 and pcl1 Show Normal Spore Viability or Meiotic Recombination in a Standard Environment
3.2. X Irradiation and Sporulation in ctt1∆ and pcl1∆ Strains
3.3. ctt1∆ Meiocytes are Highly Sensitive to ROS
3.4. Ctt1-Deficient Cells Display High Levels of ROS
3.5. Meiotic Chromosome Motility after X Irradiation
3.6. Peroxide Stress Slows Meiotic Horsetail Movements
3.7. Radical Stress Slows Prophase I Progression
3.8. IR and ROS Exposure Induces Aberrant Ascospores
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Behjati, S.; Gundem, G.; Wedge, D.C.; Roberts, N.D.; Tarpey, P.S.; Cooke, S.L.; Van Loo, P.; Alexandrov, L.B.; Ramakrishna, M.; Davies, H.; et al. Mutational signatures of ionizing radiation in second malignancies. Nat. Commun. 2016, 7, 12605. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lomax, M.E.; Folkes, L.K.; O’Neill, P. Biological consequences of radiation-induced DNA damage: Relevance to radiotherapy. Clin. Oncol. 2013, 25, 578–585. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Adewoye, A.B.; Lindsay, S.J.; Dubrova, Y.E.; Hurles, M.E. The genome-wide effects of ionizing radiation on mutation induction in the mammalian germline. Nat. Commun. 2015, 6, 6684. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Holtgrewe, M.; Knaus, A.; Hildebrand, G.; Pantel, J.-T.; Santos, M.R.d.L.; Neveling, K.; Goldmann, J.; Schubach, M.; Jäger, M.; Coutelier, M.; et al. Multisite de novo mutations in human offspring after paternal exposure to ionizing radiation. Sci. Rep. 2018, 8, 14611. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ehling, U.H.; Favor, J.; Kratochvilova, J.; Neuhäuser-Klaus, A. Dominant cataract mutations and specific-locus mutations in mice induced by radiation or ethylnitrosourea. Mutat. Res. 1982, 92, 181–192. [Google Scholar] [CrossRef]
- Illner, D.; Lorenz, A.; Scherthan, H. Meiotic chromosome mobility in fission yeast is resistant to environmental stress. Sci. Rep. 2016, 6, 24222. [Google Scholar] [CrossRef] [Green Version]
- Thorne, L.W.; Byers, B. Stage-specific effects of X-irradiation on yeast meiosis. Genetics 1993, 134, 29. [Google Scholar]
- Zickler, D.; Kleckner, N. Recombination, Pairing, and Synapsis of Homologs during Meiosis. Cold Spring Harb. Perspect. Biol. 2015, 7, a016626. [Google Scholar] [CrossRef] [Green Version]
- Adelfalk, C.; Ahmed, E.; Scherthan, H. Reproductive Phenotypes of Mouse Models Illuminate Human Infertility. J. Reprod. Med. Endocrinol. 2011, 8, 376–383. [Google Scholar]
- Saito, T.T.; Colaiacovo, M.P. Regulation of Crossover Frequency and Distribution during Meiotic Recombination. Cold Spring Harb. Symp. Quant. Biol. 2017, 82, 223–234. [Google Scholar] [CrossRef]
- Koszul, R.; Kleckner, N. Dynamic chromosome movements during meiosis: A way to eliminate unwanted connections? Trends Cell Biol. 2009, 19, 716–724. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Scherthan, H. Telomere attachment and clustering during meiosis. Cell. Mol. Life Sci. 2007, 64, 117–124. [Google Scholar] [CrossRef] [PubMed]
- Wang, P.; Zhou, R.H.; Zou, Y.; Jackson-Cook, C.K.; Povirk, L.F. Highly conservative reciprocal translocations formed by apparent joining of exchanged DNA double-strand break ends. Proc. Natl. Acad. Sci. USA 1997, 94, 12018–12023. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Iliakis, G.; Murmann, T.; Soni, A. Alternative end-joining repair pathways are the ultimate backup for abrogated classical non-homologous end-joining and homologous recombination repair: Implications for the formation of chromosome translocations. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2015, 793, 166–175. [Google Scholar] [CrossRef] [PubMed]
- Tucker, J.D. Sensitivity, specificity, and persistence of chromosome translocations for radiation biodosimetry. Mil. Med. 2002, 167, 8–9. [Google Scholar] [CrossRef] [Green Version]
- Anderson, R.M. Cytogenetic Biomarkers of Radiation Exposure. Clin. Oncol. 2019, 31, 311–318. [Google Scholar] [CrossRef] [Green Version]
- Ding, D.Q.; Chikashige, Y.; Haraguchi, T.; Hiraoka, Y. Oscillatory nuclear movement in fission yeast meiotic prophase is driven by astral microtubules, as revealed by continuous observation of chromosomes and microtubules in living cells. J. Cell Sci. 1998, 111, 701. [Google Scholar]
- Tepperberg, J.H.; Moses, M.J.; Nath, J. Colchicine effects on meiosis in the male mouse. I. Meiotic prophase: Synaptic arrest, univalents, loss of damaged spermatocytes and a possible checkpoint at pachytene. Chromosoma 1997, 106, 183–192. [Google Scholar] [CrossRef]
- Shibuya, H.; Morimoto, A.; Watanabe, Y. The dissection of meiotic chromosome movement in mice using an in vivo electroporation technique. PLoS Genet. 2014, 10, e1004821. [Google Scholar] [CrossRef] [Green Version]
- Trelles-Sticken, E.; Adelfalk, C.; Loidl, J.; Scherthan, H. Meiotic telomere clustering requires actin for its formation and cohesin for its resolution. J. Cell Biol. 2005, 170, 213–223. [Google Scholar] [CrossRef]
- Amberg, D.; Leadsham, J.E.; Kotiadis, V.; Gourlay, C.W. Cellular ageing and the actin cytoskeleton. Subcell. Biochem. 2012, 57, 331–352. [Google Scholar] [CrossRef]
- Illner, D.; Scherthan, H. Ionizing irradiation-induced radical stress stalls live meiotic chromosome movements by altering the actin cytoskeleton. Proc. Natl. Acad. Sci. USA 2013, 110, 16027. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Vogel, S.K.; Pavin, N.; Maghelli, N.; Julicher, F.; Tolic-Norrelykke, I.M. Self-organization of dynein motors generates meiotic nuclear oscillations. PLoS Biol. 2009, 7, e1000087. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Aguirre, J.; Rios-Momberg, M.; Hewitt, D.; Hansberg, W. Reactive oxygen species and development in microbiol eukaryotes. Trends Microbiol. 2005, 13, 111–118. [Google Scholar] [CrossRef] [PubMed]
- Farhood, B.; Ashrafizadeh, M.; Khodamoradi, E.; Hoseini-Ghahfarokhi, M.; Afrashi, S.; Musa, A.E.; Najafi, M. Targeting of cellular redox metabolism for mitigation of radiation injury. Life Sci. 2020, 250, 117570. [Google Scholar] [CrossRef] [PubMed]
- Culotta, V.C.; Daly, M.J. Manganese complexes: Diverse metabolic routes to oxidative stress resistance in prokaryotes and yeast. Antioxid. Redox Signal. 2013, 19, 933–944. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sharma, A.; Gaidamakova, E.K.; Grichenko, O.; Matrosova, V.Y.; Hoeke, V.; Klimenkova, P.; Conze, I.H.; Volpe, R.P.; Tkavc, R.; Gostincar, C.; et al. Across the tree of life, radiation resistance is governed by antioxidant Mn(2+), gauged by paramagnetic resonance. Proc. Natl. Acad. Sci. USA 2017, 114, E9253–E9260. [Google Scholar] [CrossRef] [Green Version]
- Chang, E.C.; Kosman, D.J. Intracellular Mn (II)-associated superoxide scavenging activity protects Cu,Zn superoxide dismutase-deficient Saccharomyces cerevisiae against dioxygen stress. J. Biol. Chem. 1989, 264, 12172–12178. [Google Scholar]
- Papadakis, M.A.; Workman, C.T. Oxidative stress response pathways: Fission yeast as archetype. Crit. Rev. Microbiol. 2015, 41, 520–535. [Google Scholar] [CrossRef] [Green Version]
- Garcia-Santamarina, S.; Boronat, S.; Calvo, I.A.; Rodriguez-Gabriel, M.; Ayte, J.; Molina, H.; Hidalgo, E. Is oxidized thioredoxin a major trigger for cysteine oxidation? Clues from a redox proteomics approach. Antioxid. Redox Signal. 2013, 18, 1549–1556. [Google Scholar] [CrossRef]
- Vivancos, A.P.; Jara, M.; Zuin, A.; Sansó, M.; Hidalgo, E. Oxidative stress in Schizosaccharomyces pombe: Different H2O2 levels, different response pathways. Mol. Genet. Genom. 2006, 276, 495–502. [Google Scholar] [CrossRef] [PubMed]
- Chen, D.; Wilkinson, C.R.; Watt, S.; Penkett, C.J.; Toone, W.M.; Jones, N.; Bahler, J. Multiple pathways differentially regulate global oxidative stress responses in fission yeast. Mol. Biol. Cell 2008, 19, 308–317. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mutoh, N.; Nakagawa, C.W.; Yamada, K. The role of catalase in hydrogen peroxide resistance in fission yeast Schizosaccharomyces pombe. Can. J. Microbiol. 1999, 45, 125–129. [Google Scholar] [CrossRef]
- Nishimoto, T.; Furuta, M.; Kataoka, M.; Kishida, M. Important role of catalase in the cellular response of the budding yeast Saccharomyces cerevisiae exposed to ionizing radiation. Curr. Microbiol. 2015, 70, 404–407. [Google Scholar] [CrossRef]
- Lorenz, A.; Osman, F.; Sun, W.; Nandi, S.; Steinacher, R.; Whitby, M.C. The fission yeast FANCM ortholog directs non-crossover recombination during meiosis. Science 2012, 336, 1585–1588. [Google Scholar] [CrossRef] [Green Version]
- Lorenz, A.; West, S.C.; Whitby, M.C. The human Holliday junction resolvase GEN1 rescues the meiotic phenotype of a Schizosaccharomyces pombe mus81 mutant. Nucleic Acids Res. 2010, 38, 1866–1873. [Google Scholar] [CrossRef] [Green Version]
- Lorenz, A.; Mehats, A.; Osman, F.; Whitby, M.C. Rad51/Dmc1 paralogs and mediators oppose DNA helicases to limit hybrid DNA formation and promote crossovers during meiotic recombination. Nucleic Acids Res. 2014, 42, 13723–13735. [Google Scholar] [CrossRef] [Green Version]
- Li, D.; Roca, M.; Yuecel, R.; Lorenz, A. Immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis. Chromosoma 2019, 128, 385–396. [Google Scholar] [CrossRef] [Green Version]
- Goldstein, A.L.; McCusker, J.H. Three new dominant drug resistance cassettes for gene disruption in Saccharomyces cerevisiae. Yeast 1999, 15, 1541–1553. [Google Scholar] [CrossRef]
- Brown, S.D.; Lorenz, A. Single-step Marker Switching in Schizosaccharomyces pombe Using a Lithium Acetate Transformation Protocol. Bio-Protocol 2016, 6, e2075. [Google Scholar] [CrossRef]
- Loidl, J.; Lorenz, A. Analysis of Schizosaccharomyces pombe Meiosis by Nuclear Spreading. Methods Mol. Biol. 2009, 558, 15–36. [Google Scholar] [CrossRef] [PubMed]
- Scherthan, H.; Adelfalk, C. Live Cell Imaging of Meiotic Chromosome Dynamics in Yeast. Methods Mol. Biol. 2011, 745, 537–548. [Google Scholar] [CrossRef] [PubMed]
- Brown, S.D.; Mpaulo, S.J.; Asogwa, M.N.; Jezequel, M.; Whitby, M.C.; Lorenz, A. DNA sequence differences are determinants of meiotic recombination outcome. Sci. Rep. 2019, 9, 16446. [Google Scholar] [CrossRef] [Green Version]
- Paulo, E.; Garcia-Santamarina, S.; Calvo, I.A.; Carmona, M.; Boronat, S.; Domenech, A.; Ayte, J.; Hidalgo, E. A genetic approach to study H2O2 scavenging in fission yeast--distinct roles of peroxiredoxin and catalase. Mol. Microbiol. 2014, 92, 246–257. [Google Scholar] [CrossRef] [Green Version]
- Dikalov, S.I.; Harrison, D.G. Methods for detection of mitochondrial and cellular reactive oxygen species. Antioxid. Redox Signal. 2014, 20, 372–382. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zielonka, J.; Kalyanaraman, B. Hydroethidine- and MitoSOX-derived red fluorescence is not a reliable indicator of intracellular superoxide formation: Another inconvenient truth. Free Radic. Biol. Med. 2010, 48, 983–1001. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wardman, P. Methods to measure the reactivity of peroxynitrite-derived oxidants toward reduced fluoresceins and rhodamines. Methods Enzymol. 2008, 441, 261–282. [Google Scholar] [CrossRef]
- Chikashige, Y.; Ding, D.Q.; Funabiki, H.; Haraguchi, T.; Mashiko, S.; Yanagida, M.; Hiraoka, Y. Telomere-led premeiotic chromosome movement in fission yeast. Science 1994, 264, 270–273. [Google Scholar] [CrossRef]
- Watanabe, Y.; Nurse, P. Cohesin Rec8 is required for reductional chromosome segregation at meiosis. Nature 1999, 400, 461–464. [Google Scholar] [CrossRef]
- Sato, M.; Toda, T. Alp7/TACC is a crucial target in Ran-GTPase-dependent spindle formation in fission yeast. Nature 2007, 447, 334–337. [Google Scholar] [CrossRef]
- Lorenz, A.; Estreicher, A.; Kohli, J.; Loidl, J. Meiotic recombination proteins localize to linear elements in Schizosaccharomyces pombe. Chromosoma 2006, 115, 330–340. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Azzam, E.I.; Jay-Gerin, J.-P.; Pain, D. Ionizing radiation-induced metabolic oxidative stress and prolonged cell injury. Cancer Lett. 2012, 327, 48–60. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Nishimoto, T.; Watanabe, T.; Furuta, M.; Kataoka, M.; Kishida, M. Roles of Catalase and Trehalose in the Protection from Hydrogen Peroxide Toxicity in Saccharomyces cerevisiae. Biocontrol Sci. 2016, 21, 179–182. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Le Caër, S. Water Radiolysis: Influence of Oxide Surfaces on H2 Production under Ionizing Radiation. Water 2011, 3, 235–253. [Google Scholar] [CrossRef] [Green Version]
- Aguirre, J.; Hansberg, W.; Navarro, R. Fungal responses to reactive oxygen species. Med. Mycol. 2006, 44, S101–S107. [Google Scholar] [CrossRef] [Green Version]
Strain Name | Genotype | Origin/Source |
---|---|---|
ALP714 | h+S | [35] |
ALP731 | h-smt0 ade6-469 his3+-aim arg3-D4 his3-D1 ura4-D18 | [36] |
ALP733 | h+S ade6-3083 ura4+-aim2 his3-D1 leu1-32 ura4-D18 | [36] |
ALP1596 | h-smt0 ade7-152 his3-D1 leu1-32 ura4-D18 | [37] |
FO652 | h-smt0 arg3-D4 his3-D1 leu1-32 ura4-D18 | [37] |
UoA794 a | h+S GFP-atb2+::natMX4 rec8+::GFP-kanMX6 ade6-M210 his1-102 leu1-32 | this study |
UoA795 a | h-smt0 GFP-atb2+::natMX4 rec8+::GFP-kanMX6 ade6-M216 leu2-120 his7-366 | this study |
UoA798 (WT) | h+S/h-smt0 GFP-atb2+::natMX4/GFP-atb2+::natMX4 rec8+::GFP-kanMX6/rec8+::GFP-kanMX6 ade6-M210/ade6-M216 his1-102/his1+ leu2-120/leu2+ leu1-32/leu1+ his7-366/his7+ | this study; cross of UoA794 × UoA795 |
UoA992 | h+S pcl1∆-21a::hphMX4 GFP-atb2+::natMX4 rec8+::GFP-kanMX6 ade6-M210 his1-102 leu1-32 | this study; derivative of UoA794 |
UoA993 | h-smt0 pcl1∆-21b::hphMX4 GFP-atb2+::natMX4 rec8+::GFP-kanMX6 ade6-M216 leu2-120 his7-366 | this study; derivative of UoA795 |
UoA994 (pcl1∆) | h+S/h-smt0 pcl1∆-21a::hphMX4/pcl1∆-21b::hphMX4 GFP-atb2+::natMX4/GFP-atb2+::natMX4 rec8+::GFP-kanMX6/rec8+::GFP-kanMX6 ade6-M210/ade6-M216 his1-102/his1+ leu2-120/leu2+ leu1-32/leu1+ his7-366/his7+ | this study; cross of UoA992 × UoA993 |
UoA995 | h+S ctt1∆-18a::hphMX4 GFP-atb2+::natMX4 rec8+::GFP-kanMX6 ade6-M210 his1-102 leu1-32 | this study; derivative of UoA794 |
UoA996 | h-smt0 ctt1∆-18b::hphMX4 GFP-atb2+::natMX4 rec8+::GFP-kanMX6 ade6-M216 leu2-120 his7-366 | this study; derivative of UoA795 |
UoA997 (ctt1∆) | h+S/h-smt0 ctt1∆-18a::hphMX4/ctt1∆-18b::hphMX4 GFP-atb2+::natMX4/GFP-atb2+::natMX4 rec8+::GFP-kanMX6/rec8+::GFP-kanMX6 ade6-M210/ade6-M216 his1-102/his1+ leu2-120/leu2+ leu1-32/leu1+ his7-366/his7+ | this study; cross of UoA995 × UoA996 |
UoA1038 | h-smt0 pcl1∆-21c::hphMX4 arg3-D4 his3-D1 leu1-32 ura4-D18 | this study; derivative of FO652 |
UoA1039 | h-smt0 ctt1∆-18c::hphMX4 arg3-D4 his3-D1 leu1-32 ura4-D18 | this study; derivative of FO652 |
UoA1040 | h+S pcl1∆-21c::hphMX4 ade6-3083 ura4+-aim2 his3-D1 leu1-32 ura4-D18 | this study |
UoA1043 | h-smt0 pcl1∆-21c::hphMX4 ade6-469 his3+-aim arg3-D4 his3-D1 ura4-D18 | this study |
UoA1044 | h+S ctt1∆-18c::hphMX4 ade6-3083 ura4+-aim2 his3-D1 leu1-32 ura4-D18 | this study |
UoA1047 | h-smt0 ctt1∆-18c::hphMX4 ade6-469 his3+-aim arg3-D4 his3-D1 ura4-D18 | this study |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Muhtadi, R.; Lorenz, A.; Mpaulo, S.J.; Siebenwirth, C.; Scherthan, H. Catalase T-Deficient Fission Yeast Meiocytes Show Resistance to Ionizing Radiation. Antioxidants 2020, 9, 881. https://doi.org/10.3390/antiox9090881
Muhtadi R, Lorenz A, Mpaulo SJ, Siebenwirth C, Scherthan H. Catalase T-Deficient Fission Yeast Meiocytes Show Resistance to Ionizing Radiation. Antioxidants. 2020; 9(9):881. https://doi.org/10.3390/antiox9090881
Chicago/Turabian StyleMuhtadi, Razan, Alexander Lorenz, Samantha J. Mpaulo, Christian Siebenwirth, and Harry Scherthan. 2020. "Catalase T-Deficient Fission Yeast Meiocytes Show Resistance to Ionizing Radiation" Antioxidants 9, no. 9: 881. https://doi.org/10.3390/antiox9090881