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MBC in Press, published online ahead of print November 24, 2004
Mol. Biol. Cell 10.1091/mbc.E04-09-0842

A more recent version of this article appeared on February 1, 2005
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Submitted on September 27, 2004
Accepted on November 5, 2004

A Sorting Nexin PpAtg24 Regulates Vacuolar Membrane Dynamics during Pexophagy via Binding to Phosphatidylinositol-3-Phosphate

Yoshitaka Ano,* Takeshi Hattori,* Masahide Oku,* Hiroyuki Mukaiyama,* Misuzu Baba,{dagger} Yoshinori Ohsumi,{ddagger} Nobuo Kato,* and Yasuyoshi Sakai*{ddagger}{sect}

*Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan; {dagger}Department of Chemical and Biological Sciences, Faculty of Science, Japan Women’s University, Tokyo 112-8681, Japan; {ddagger}Department of Cell Biology, National Institute for Basic Biology, Okazaki 444-8585, Japan

Monitoring Editor: Howard Riezman

Diverse cellular processes such as autophagic protein degradation require phosphoinositide signaling in eukaryotic cells. In the methylotrophic yeast Pichia pastoris, peroxisomes can be selectively degraded via two types of pexophagic pathways, macropexophagy and micropexophagy. Both involve membrane fusion events at the vacuolar surface that are characterized by internalization of the boundary domain of the fusion complex, indicating that fusion occurs at the vertex. Here, we show that PpAtg24, a molecule with a phosphatidylinositol 3-phosphate-binding module (PX domain) that is indispensible for pexophagy, functions in membrane fusion at the vacuolar surface. CFP-tagged PpAtg24 localized to the vertex and boundary region of the pexophagosome-vacuole fusion complex during macropexophagy. Depletion of PpAtg24 resulted in the blockage of macropexophagy after pexophagosome formation and before the fusion stage. These and other results suggest that PpAtg24 is involved in the spatiotemporal regulation of membrane fusion at the vacuolar surface during pexophagy via binding to phosphatidyl inositol 3-phosphate, rather than the previously suggested function in formation of the pexophagosome.

{sect}Corresponding author. E-mail: ysakai{at}kais.kyoto-u.ac.jp




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