We have identified and purified six subforms of peroxisomes, designated P1 to P6, from the yeast, Yarrowia lipolytica. An analysis of trafficking of peroxisomal proteins in vivo suggests the existence of a multistep peroxisome assembly pathway in Y. lipolytica. This pathway operates by conversion of peroxisomal subforms in the direction P1, P2→P3→P4→P5→P6 and involves the import of various peroxisomal proteins into distinct vesicular intermediates. We have also reconstituted in vitro the fusion of the earliest intermediates in the pathway, small peroxisomal vesicles P1 and P2. Their fusion leads to the formation of a larger and more dense peroxisomal vesicle, P3. Fusion of P1 and P2 in vitro requires cytosol and ATP hydrolysis and is inhibited by antibodies to two membrane-associated ATPases of the AAA family, Pex1p and Pex6p. We provide evidence that the fusion in vitro of P1 and P2 peroxisomes reconstructs an actual early step in the peroxisome assembly pathway operating in vivo in Y. lipolytica.
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10 January 2000
Article|
January 10 2000
Fusion of Small Peroxisomal Vesicles in Vitro Reconstructs an Early Step in the in Vivo Multistep Peroxisome Assembly Pathway of Yarrowia lipolytica
Vladimir I. Titorenko,
Vladimir I. Titorenko
aDepartment of Cell Biology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada
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Honey Chan,
Honey Chan
aDepartment of Cell Biology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada
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Richard A. Rachubinski
Richard A. Rachubinski
aDepartment of Cell Biology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada
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Vladimir I. Titorenko
aDepartment of Cell Biology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada
Honey Chan
aDepartment of Cell Biology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada
Richard A. Rachubinski
aDepartment of Cell Biology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada
Abbreviations used in this paper: AOX, acyl-CoA oxidase; HSP, high-speed pelletable peroxisomes; NSF, N-ethylmaleimide-sensitive factor; SNAPs, soluble NSF attachment proteins; SNAREs, SNAP receptors; v-SNAREs and t-SNAREs, vesicle- and target-specific integral membrane proteins, respectively.
Received:
September 10 1999
Revision Requested:
December 02 1999
Accepted:
December 02 1999
Online ISSN: 1540-8140
Print ISSN: 0021-9525
© 2000 The Rockefeller University Press
2000
The Rockefeller University Press
J Cell Biol (2000) 148 (1): 29–44.
Article history
Received:
September 10 1999
Revision Requested:
December 02 1999
Accepted:
December 02 1999
Citation
Vladimir I. Titorenko, Honey Chan, Richard A. Rachubinski; Fusion of Small Peroxisomal Vesicles in Vitro Reconstructs an Early Step in the in Vivo Multistep Peroxisome Assembly Pathway of Yarrowia lipolytica. J Cell Biol 10 January 2000; 148 (1): 29–44. doi: https://doi.org/10.1083/jcb.148.1.29
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