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J Biol Chem, Vol. 274, Issue 50, 35293-35296, December 10, 1999

COMMUNICATION
Formation of Peroxisomes from Peroxisomal Ghosts in a Peroxisome-deficient Mammalian Cell Mutant upon Complementation by Protein Microinjection^*

Masatoshi Yamasaki, Noriyo Hashiguchi, Chiharu Fujiwara, Tsuneo Imanaka^§, Toshiro Tsukamoto, and Takashi Osumi^¶

From the  Department of Life Science, Faculty of Science, Himeji Institute of Technology, 3-2-1 Koto, Kamigori, Hyogo 678-1297, Japan and the ^§ Laboratory of Cell Biology, Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, 2630 Sugitani, Toyama 930-0194, Japan

Most mammalian cell strains genetically deficient in peroxisome biogenesis have abnormal membrane structures called ghosts, containing integral peroxisomal membrane protein, PMP70, but lacking the peroxisomal matrix proteins. Upon genetic complementation, these mutants regain the ability of peroxisome biogenesis. It is postulated that, in this process, the ghosts act as the precursors of peroxisomes, but there has been no evidence to support this. In the present study, we investigated this issue by protein microinjection to a mutant Chinese hamster ovary cell line defective of PEX5, encoding a peroxisome-targeting signal receptor. When recombinant Pex5p and green fluorescent protein (GFP) carrying a peroxisome-targeting signal were co-injected into the mutant cells, the GFP fluorescence gathered over time to particulate structures where PMP70 was co-localized. This process was dependent on both Pex5p and the targeting signal, and, most importantly, occurred even in the presence of cycloheximide, a protein synthesis inhibitor. These findings suggest that the ghosts act as acceptors of matrix proteins in the peroxisome recovery process at least in the PEX5 mutant, and support the view that peroxisomes can grow by incorporating newly synthesized matrix proteins.

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^* This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture of Japan.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom all correspondence should be addressed. Tel.: +81-791-58-0192; Fax: +81-791-58-0193; E-mail: osumi@sci.himeji-tech.ac.jp.

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Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.

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