Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
권호 표지
DOI QR코드

DOI QR Code

Identification of Atg8 Isoform in Encysting Acanthamoeba

  • Moon, Eun-Kyung (Department of Parasitology, Kyungpook National University School of Medicine) ;
  • Hong, Yeonchul (Department of Parasitology, Kyungpook National University School of Medicine) ;
  • Chung, Dong-Il (Department of Parasitology, Kyungpook National University School of Medicine) ;
  • Kong, Hyun-Hee (Department of Parasitology, Dong-A University College of Medicine)
  • Received : 2013.06.26
  • Accepted : 2013.08.29
  • Published : 2013.10.31
Download PDF
⟨ Previous Next ⟩

Abstract

Autophagy-related protein 8 (Atg8) is an essential component of autophagy formation and encystment of cystforming parasites, and some protozoa, such as, Acanthamoeba, Entamoeba, and Dictyostelium, have been reported to possess a type of Atg8. In this study, an isoform of Atg8 was identified and characterized in Acanthamoeba castellanii (AcAtg8b). AcAtg8b protein was found to encode 132 amino acids and to be longer than AcAtg8 protein, which encoded 117 amino acids. Real-time PCR analysis showed high expression levels of AcAtg8b and AcAtg8 during encystation. Fluorescence microscopy demonstrated that AcAtg8b is involved in the formation of the autophagosomal membrane. Chemically synthesized siRNA against AcAtg8b reduced the encystation efficiency of Acanthamoeba, confirming that AcAtg8b, like AcAtg8, is an essential component of cyst formation in Acanthamoeba. Our findings suggest that Acanthamoeba has doubled the number of Atg8 gene copies to ensure the successful encystation for survival when 1 copy is lost. These 2 types of Atg8 identified in Acanthamoeba provide important information regarding autophagy formation, encystation mechanism, and survival of primitive, cyst-forming protozoan parasites.

Keywords

References

  1. Levine B, Klionsky DJ. Development by self-digestion: molecular mechanisms and biological functions of autophagy. Dev Cell 2004; 6: 463-477. https://doi.org/10.1016/S1534-5807(04)00099-1
  2. Picazarri K, Nakada-Tsukui K, Nozaki T. Autophagy during proliferation and encystation in the protozoan parasite Entamoeba invadens. Infect Immun 2008; 76: 278-288. https://doi.org/10.1128/IAI.00636-07
  3. Shpilka T, Weidberg H, Pietrokovski S, Elazar Z. Atg8: an autophagy- related ubiquitin-like protein family. Genome Biol 2011; 12: 226-236. https://doi.org/10.1186/gb-2011-12-7-226
  4. Suzuki K, Ohsumi Y. Molecular machinery of autophagosome formation in yeast, Saccharomyces cerevisiae. FEBS Lett 2007; 581: 2156-2161. https://doi.org/10.1016/j.febslet.2007.01.096
  5. Kuma A, Mizushima N, Ishihara N, Ohsumi Y. Formation of the approximately 350-kDa Apg12-Apg5.Apg16 multimeric complex, mediated by Apg16 oligomerization, is essential for autophagy in yeast. J Biol Chem 2002; 277: 18619-18625. https://doi.org/10.1074/jbc.M111889200
  6. Mizushima N, Noda T, Yoshimori T, Tanaka Y, Ishii T, George MD, Klionsky DJ, Ohsumi M, Ohsumi Y. A protein conjugation system essential for autophagy. Nature 1998; 395: 395-398. https://doi.org/10.1038/26506
  7. Moon EK, Chung DI, Hong YC, Kong HH. Autophagy protein 8 mediating autophagosome in encysting Acanthamoeba. Mol Biochem Parasitol 2009; 168: 43-48. https://doi.org/10.1016/j.molbiopara.2009.06.005
  8. Song SM, Han BI, Moon EK, Lee YR, Yu HS, Jha BK, Danne DB, Kong HH, Chung DI, Hong Y. Autophagy protein 16-mediated autophagy is required for the encystation of Acanthamoeba castellanii. Mol Biochem Parasitol 2012; 183: 158-165. https://doi.org/10.1016/j.molbiopara.2012.02.013
  9. Lee JY, Song SM, Moon EK, Lee YR, Jha BK, Danne DB, Cha HJ, Yu HS, Kong HH, Chung DI, Hong Y. Cysteine Protease Inhibitor (AcStefin) Is Required for Complete Cyst Formation of Acanthamoeba. Eukaryot Cell. 2013; 12: 567-574. https://doi.org/10.1128/EC.00308-12
  10. Moon EK, Chung DI, Hong YC, Kong HH. Characterization of a serine proteinase mediating encystation of Acanthamoeba. Eukaryot Cell 2008; 7: 1513-1517. https://doi.org/10.1128/EC.00068-08
  11. Moon EK, Chung DI, Hong Y, Kong HH. Atg3-mediated lipidation of Atg8 is involved in encystation of Acanthamoeba. Korean J Parasitol 2011; 49: 103-108. https://doi.org/10.3347/kjp.2011.49.2.103
  12. Moon EK, Hong Y, Chung DI, Kong HH. Cysteine protease involving in autophagosomal degradation of mitochondria during encystation of Acanthamoeba. Mol Biochem Parasitol 2012; 185: 121-126. https://doi.org/10.1016/j.molbiopara.2012.07.008
  13. Eichinger L, Pachebat JA, Glöckner G, Rajandream MA, Sucgang R, Berriman M, Song J, Olsen R, Szafranski K, Xu Q, Tunggal B, Kummerfeld S, Madera M, Konfortov BA, Rivero F, Bankier AT, Lehmann R, Hamlin N, Davies R, Gaudet P, Fey P, Pilcher K, Chen G, Saunders D, Sodergren E, Davis P, Kerhornou A, Nie X, Hall N, Anjard C, Hemphill L, Bason N, Farbrother P, Desany B, Just E, Morio T, Rost R, Churcher C, Cooper J, Haydock S, van Driessche N, Cronin A, Goodhead I, Muzny D, Mourier T, Pain A, Lu M, Harper D, Lindsay R, Hauser H, James K, Quiles M, Madan Babu M, Saito T, Buchrieser C, Wardroper A, Felder M, Thangavelu M, Johnson D, Knights A, Loulseged H, Mungall K, Oliver K, Price C, Quail MA, Urushihara H, Hernandez J, Rabbinowitsch E, Steffen D, Sanders M, Ma J, Kohara Y, Sharp S, Simmonds M, Spiegler S, Tivey A, Sugano S, White B, Walker D, Woodward J, Winckler T, Tanaka Y, Shaulsky G, Schleicher M, Weinstock G, Rosenthal A, Cox EC, Chisholm RL, Gibbs R, Loomis WF, Platzer M, Kay RR, Williams J, Dear PH, Noegel AA, Barrell B, Kuspa A. The genome of the social amoeba Dictyostelium discoideum. Nature 2005; 435: 43-57. https://doi.org/10.1038/nature03481
  14. Kong HH, Pollard TD. Intracellular localization and dynamics of myosin-II and myosin-IC in live Acanthamoeba by transient transfection of EGFP fusion proteins. J Cell Sci 2002; 15: 4993-5002.
  15. Bowers B, Korn ED. The fine structure of Acanthamoeba castellanii (Neff strain). II. Encystment. J Cell Biol 1969; 41: 786-805. https://doi.org/10.1083/jcb.41.3.786
  16. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)). Method Methods 2001; 25: 402-408. https://doi.org/10.1006/meth.2001.1262
  17. Bagchi S, Oniku AE, Topping K, Mamhoud ZN, Paget TA. Programmed cell death in Giardia. Parasitology 2012; 139: 894-903. https://doi.org/10.1017/S003118201200011X
  18. Kanki T, Wang K, Cao Y, Baba M, Klionsky DJ. Atg32 is a mitochondrial protein that confers selectivity during mitophagy. Dev Cell 2009; 17: 98-109. https://doi.org/10.1016/j.devcel.2009.06.014
  19. Kissová I, Deffieu M, Manon S, Camougrand N. Uth1p is involved in the autophagic degradation of mitochondria. J Biol Chem 2004; 279: 39068-39074. https://doi.org/10.1074/jbc.M406960200
  20. Tal R, Winter G, Ecker N, Klionsky DJ, Abeliovich H. Aup1p, a yeast mitochondrial protein phosphatase homolog, is required for efficient stationary phase mitophagy and cell survival. J Biol Chem 2007; 282: 5617-5624. https://doi.org/10.1074/jbc.M605940200

Cited by

  1. Autophagy protein 12 plays an essential role in Acanthamoeba encystation vol.159, pp.None, 2013, https://doi.org/10.1016/j.exppara.2015.08.013
  2. Encystation: the most prevalent and underinvestigated differentiation pathway of eukaryotes vol.164, pp.5, 2013, https://doi.org/10.1099/mic.0.000653
  3. Ubiquitin-Like Modifiers: Emerging Regulators of Protozoan Parasites vol.10, pp.10, 2013, https://doi.org/10.3390/biom10101403
  4. Peganum harmala Extract Has Antiamoebic Activity to Acanthamoeba triangularis Trophozoites and Changes Expression of Autophagy-Related Genes vol.10, pp.7, 2021, https://doi.org/10.3390/pathogens10070842
  5. Amoebicidal activity of Cassia angustifolia extract and its effect on Acanthamoeba triangularis autophagy-related gene expression at the transcriptional level vol.148, pp.9, 2013, https://doi.org/10.1017/s0031182021000718