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MBC in Press, published online ahead of print December 7, 2002
Mol. Biol. Cell 10.1091/mbc.E02-08-0539

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Submitted on August 27, 2002
Revised on October 14, 2002
Accepted on October 21, 2002

Pex7p and Pex20p of Neurospora crassa function together in PTS2-dependent protein import into peroxisomes

Martin Sichting1, Annette Schell-Steven2, Holger Prokisch3, Ralf Erdmann2, and Hanspeter Rottensteiner2*

1 Freie Universität Berlin, Fachbereich Biologie, Chemie, Pharmazie, Thielallee 63, D-14195 Berlin, Germany (present address: Ludwig Maximilian Universität München, Butenandtstrasse 5, Haus B, D-81377 München, Germany)
2 Freie Universität Berlin, Fachbereich Biologie, Chemie, Pharmazie, Thielallee 63, D-14195 Berlin, Germany
3 Ludwig Maximilian Universität München, Butenandtstrasse 5, Haus B, D-81377 München, Germany

* Corresponding author. E-mail address: hpr{at}zedat.fu-berlin.de.

Recruiting matrix proteins with a peroxisomal targeting signal type 2 (PTS2) to the peroxisomal membrane requires species-specific factors. In Saccharomyces cerevisiae, the PTS2 receptor Pex7p acts in concert with the redundant Pex18p/Pex21p, while in Yarrowia lipolytica, Pex20p might unite the function of both S. cerevisiae peroxins. Here, the genome of the filamentous fungus Neurospora crassa was analyzed for peroxin-encoding genes. We identified a set of 18 peroxins that resembles that of Y. lipolytica rather than that of S. cerevisiae. Interestingly, proteins homologous to both S. cerevisiae Pex7p and Y. lipolytica Pex20p exist in N. crassa. We report on the isolation of these PTS2-specific peroxins and demonstrate that NcPex20p can substitute for S. cerevisiae Pex18p/Pex21p, but not for ScPex7p. Like Pex18p, NcPex20p did not bind PTS2 protein or the docking proteins in the absence of ScPex7p. Rather, NcPex20p was required before docking to form an import-competent complex of cargo-loaded PTS2 receptors. NcPex7p did not functionally replace yeast Pex7p, probably because the N. crassa PTS2 receptor failed to associate with Pex18p/Pex21p. However, once NcPex7p and NcPex20p had been co-expressed, it proved possible to replace yeast Pex7p. Pex20p and Pex18p/Pex21p are therefore true orthologues, both of which are in need of Pex7p for PTS2 protein import.




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