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Published 3 January 2006. doi:10.1083/jcb.200508096
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 172, Number 1, 67-78
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Article

Dynamics of the peroxisomal import cycle of PpPex20p : ubiquitin-dependent localization and regulation



Sébastien Léon1, Lan Zhang1, W. Hayes McDonald2, John Yates, III2, James M. Cregg3, and Suresh Subramani1

1 Section of Molecular Biology, Division of Biological Sciences, University of California, San Diego, La Jolla, CA 92093
2 The Scripps Research Institute, San Diego, CA 92037
3 Keck Graduate Institute, Claremont, CA 91711

Correspondence to Suresh Subramani: ssubramani{at}ucsd.edu

We characterize the peroxin PpPex20p from Pichia pastoris and show its requirement for translocation of PTS2 cargoes into peroxisomes. PpPex20p docks at the peroxisomal membrane and translocates into peroxisomes. Its peroxisomal localization requires the docking peroxin Pex14p but not the peroxins Pex2p, Pex10p, and Pex12p, whose absence causes peroxisomal accumulation of Pex20p. Similarities between Pex5p and Pex20p were noted in their protein interactions and dynamics during import, and both contain a conserved NH2-terminal domain. In the absence of the E2-like Pex4p or the AAA proteins Pex1p and Pex6p, Pex20p is degraded via polyubiquitylation of residue K19, and the K19R mutation causes accumulation of Pex20p in peroxisome remnants. Finally, either interference with K48-branched polyubiquitylation or removal of the conserved NH2-terminal domain causes accumulation of Pex20p in peroxisomes, mimicking a defect in its recycling to the cytosol. Our data are consistent with a model in which Pex20p enters peroxisomes and recycles back to the cytosol in an ubiquitin-dependent manner.

Abbreviations used in this paper: 3-AT, 3-aminotriazole; BFP, blue fluorescent protein; DIC, differential interference contrast; mPTS, membrane PTS; mRFP, monomeric red fluorescent protein; PNS, postnuclear supernatant; PTS, peroxisomal targeting signal; RADAR, receptor accumulation and degradation in the absence of recycling; RING, really interesting new gene; UPS, ubiquitin–proteasome system.


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