Autophagosome formation can be achieved in the absence of Atg18 by expressing engineered PAS-targeted Atg2.
The Atg2-Atg18 complex is essential for autophagosome formation in Saccharomyces cerevisiae. In this paper, we show that partial induction of autophagy can proceed in cells expressing engineered variants of Atg2 capable of localizing to the pre-autophagosomal structure (PAS) in the absence of Atg18. Specifically, through the construction of fusion proteins, we show that the fusion to Atg2 of either the phosphatidylinositol 3-phosphate-binding FYVE domain or the core autophagy protein Atg8 allowed limited Atg18-independent recovery of autophagosome formation. These results indicate that effective targeting of Atg2 to the PAS can compensate for loss of Atg18 function in autophagy.
Frontier Research Center, Tokyo Institute of Technology, Yokohama 226-8503, Japan.
Takafumi
T
Kobayashi
Kuninori
K
Suzuki
Yoshinori
Y
Ohsumi
D001343:Autophagy; D000071183:Autophagy-Related Proteins; D003593:Cytoplasm / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005818:Genetic Engineering; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D008854:Microscopy, Electron / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D009154:Mutation; D018129:Phosphatidylinositol Phosphates / Q000737:chemistry; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*
C441008:ATG18 protein, S cerevisiae; C434064:ATG2 protein, S cerevisiae; D000071183:Autophagy-Related Proteins; D008565:Membrane Proteins; D018129:Phosphatidylinositol Phosphates; D029701:Saccharomyces cerevisiae Proteins; C055525:phosphatidylinositol 3-phosphate; D049452:Green Fluorescent Proteins