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MBC in Press, published online ahead of print September 17, 2003
Mol. Biol. Cell 10.1091/mbc.E03-05-0340

A more recent version of this article appeared on January 1, 2004
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Submitted on May 28, 2003
Revised on August 17, 2003
Accepted on August 22, 2003

Modification of a Ubiquitin-like Protein Paz2 Conducted Micropexophagy through Formation of a Novel Membrane Structure

Hiroyuki Mukaiyama1, Misuzu Baba2, Masako Osumi3, Satoshi Aoyagi1, Nobuo Kato1, Yoshinori Ohsumi4, and Yasuyoshi Sakai5*

1 Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto 606-8502, Japan
2 Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University, Mejirodai, Bunkyo-ku, Tokyo 112-8681, Japan; Department of Cell Biology, National Institute for Basic Biology, Okazaki, 444-8585, Japan
3 Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University, Mejirodai, Bunkyo-ku, Tokyo 112-8681, Japan
4 Department of Cell Biology, National Institute for Basic Biology, Okazaki, 444-8585, Japan
5 Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto 606-8502, Japan, Department of Cell Biology, National Institute for Basic Biology, Okazaki, 444-8585, Japan

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

Microautophagy is a versatile process in which vacuolar or lysosomal membranes directly sequester cytosolic targets for degradation. Recent genetic evidence suggested that microautophagy utilizes molecular machineries essential for macroautophagy, but the details of this process are still unknown. In this study, a ubiquitin-like protein Paz2 essential for micropexophagy in the yeast Pichia pastoris has been shown to receive modification through the function of Paz8 and Gsa7, yielding a modified form Paz2-I, similar to the ubiquitin-like lipidation of Aut7 that is essential for macroautophagy in S. cerevisiae. We identified a novel membrane structure formed after the onset of micropexophagy, which we suggest is necessary for the sequestration of peroxisomes by the vacuole. Assembly of this newly formed membrane structure, which is followed by localization of Paz2 to it, was found to require a properly functioning Paz2-modification system. We herein show that Paz2 and its modification system conduct micropexophagy through formation of the membrane structure, which explains the convergence between micropexophagy and macroautophagy with regards to de novo membrane formation.




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