Characterization of the peroxiredoxin 1 subfamily from Tetrahymena thermophila
- 124 Downloads
Peroxiredoxins are antioxidant enzymes that use redox active Cys residues to reduce H2O2 and various organic hydroperoxides to less reactive products, and thereby protect cells against oxidative stress. In yeasts and mammals, the Prx1 proteins are sensitive to hyperoxidation and consequent loss of their peroxidase activity whereas in most bacteria they are not. In this paper we report the characterization of the Prx1 family in the non-parasitic protist Tetrahymena thermophila. In this organism, four genes potentially encoding Prx1 have been identified. In particular, we show that the mitochondrial Prx1 protein (Prx1m) from T. thermophila is relatively robust to hyperoxidation. This is surprising given that T. thermophila is a eukaryote like yeasts and mammals. In addition, the proliferation of the T. thermophila cells was relatively robust to inhibition by H2O2, cumene hydroperoxide and plant natural products that are known to promote the production of H2O2. In the presence of these agents, the abundance of the T. thermophila Prx1m protein was shown to increase. This suggested that the Prx1m protein may be protecting the cells against oxidative stress. There was no evidence for any increase in Prx1m gene expression in the stressed cells. Thus, increasing protein stability rather than increasing gene expression may explain the increasing Prx1m protein abundance we observed.
KeywordsAlveolates Ciliated protozoa Antioxidant enzymes Oxidative stress Plant natural products
Funding was provided by Flinders University and a PhD scholarship awarded to Sarmad Al-Asadi by the Government of Iraq.
- 5.Biteau B, Labarre J, Toledano MB (2003) ATP-dependent reduction of cysteine-sulphinic acid by S. cerevisiae sulphiredoxin. Nature 425(6961):980–984Google Scholar
- 7.Rhee SG, Jeong W, Chang TS, Woo HA (2007) Sulfiredoxin, the cysteine sulfinic acid reductase specific to 2-Cys peroxiredoxin: its discovery, mechanism of action, and biological significance. Kidney Int 72:S3–S8Google Scholar
- 13.Cassidy-Hanley DM (2012) Tetrahymena in the laboratory: strain resources, methods for culture, maintenance, and storage. In: Collins K (ed) Tetrahymena thermophila. Elsevier Academic Press Inc, San Diego, pp 239–276Google Scholar
- 15.Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4):402–408Google Scholar
- 16.Tikhonenkov DV, Janouskovec J, Mylnikov AP, Mikhailov KV, Simdyanov TG, Aleoshin VV, Keeling PJ (2014) Description of Colponema vietnamica sp.n. and Acavomonas peruviana n. gen. n. sp., two new alveolate phyla (Colponemidia nom. nov and Acavomonidia nom. nov.) and their contributions to reconstructing the ancestral state of alveolates and eukaryotes. PLoS One 9(4):e95467PubMedPubMedCentralGoogle Scholar
- 17.Woo YH, Ansari H, Otto TD, Klinger CM, Kolisko M, Michalek J, Saxena A, Shanmugam D, Tayyrov A, Veluchamy A, Ali S, Bernal A, del Campo J, Cihlar J, Flegontov P, Gornik SG, Hajduskova E, Hora A, Janouskovec J, Katris NJ, Mast FD, Miranda-Saavedra D, Mourier T, Naeem R, Nair M, Panigrahi AK, Rawlings ND, Padron-Regalado E, Ramaprasad A, Samad N, Tomcala A, Wilkes J, Neafsey DE, Doerig C, Bowler C, Keeling PJ, Roos DS, Dacks JB, Templeton TJ, Waller RF, Lukes J, Obornik M, Pain A (2015) Chromerid genomes reveal the evolutionary path from photosynthetic algae to obligate intracellular parasites. Elife 4:e06974PubMedPubMedCentralGoogle Scholar
- 23.Goncalves I, Duret L, Mouchiroud D (2000) Nature and structure of human genes that generate retropseudogenes. Genome 10(5):672–678Google Scholar
- 25.Shang Y, Song X, Bowen J, Corstanje R, Gao Y, Gaertig J, Gorovsky MA (2002) A robust inducible-repressible promoter greatly facilitates gene knockouts, conditional expression, and overexpression of homologous and heterologous genes in Tetrahymena thermophila. Proc Natl Acad Sci USA 99(6):3734–3739PubMedGoogle Scholar
- 40.Wu XJ, Kassie F, Mersch-Sundermann V (2005) Induction of apoptosis in tumor cells by naturally occurring sulfur-containing compounds. Mutat Res Rev Mutat Res 589(2):81–102Google Scholar
- 42.Deponte M (2008) Programmed cell death in protists. Biochim Biophys Acta Mol Cell Res 1783(7):1396–1405Google Scholar
- 45.Jang HH, Lee KO, Chi YH, Jung BG, Park SK, Park JH, Lee JR, Lee SS, Moon JC, Yun JW, Choi YO, Kim WY, Kang JS, Cheong GW, Yun DJ, Rhee SG, Cho MJ, Lee SY (2004) Two enzymes in one: two yeast peroxiredoxins display oxidative stress-dependent switching from a peroxidase to a molecular chaperone function. Cell 117(5):625–635PubMedGoogle Scholar