Korean J Parasitol.  2011 Jun;49(2):103-108. 10.3347/kjp.2011.49.2.103.

Atg3-Mediated Lipidation of Atg8 Is Involved in Encystation of Acanthamoeba

• Moon EK
• Chung DI
• Hong Y
• Kong HH

• KMID: 1090608
• DOI: http://doi.org/10.3347/kjp.2011.49.2.103

Abstract

Autophagy is a catabolic process involved in the degradation of a cell's own components for cell growth, development, homeostasis, and the recycling of cellular products. Autophagosome is an essential component in the protozoan parasite during differentiation and encystation. The present study identified and characterized autophagy-related protein (Atg) 3, a member of Atg8 conjugation system, in Acanthamoeba castellanii (AcAtg3). AcAtg3 encoding a 304 amino acid protein showed high similarity with the catalytic cysteine site of other E2 like enzymes of ubiquitin system. Predicted 3D structure of AcAtg3 revealed a hammer-like shape, which is the characteristic structure of E2-like enzymes. The expression level of AcAtg3 did not increase during encystation. However, the formation of mature cysts was significantly reduced in Atg3-siRNA transfected cells in which the production of Atg8-phosphatidylethanolamine conjugate was inhibited. Fluorescent microscopic analysis revealed that dispersed AcAtg3-EGFP fusion protein gathered around autophagosomal membranes during encystation. These results provide important information for understanding autophagic machinery through the lipidation reaction mediated by Atg3 in Acanthamoeba.

Keyword

Acanthamoeba; encystation; autophagosome; Atg3

MeSH Terms

Acanthamoeba castellanii/*growth & development/*metabolism
Animals
Gene Knockdown Techniques
Lipid Metabolism
Models, Molecular
Molecular Sequence Data
Protein Structure, Tertiary
Protozoan Proteins/genetics
RNA, Small Interfering/metabolism
Rats
Sequence Analysis, DNA
Spores, Protozoan/*growth & development/*metabolism
Ubiquitin-Conjugating Enzymes/genetics/*metabolism