title: Overproduction of Pex5p stimulates import of alcohol oxidase and dihydroxyacetone synthase in a Hansenula polymorpha Pex14 null mutant.
abstract: Hansenula polymorpha Deltapex14 cells are affected in peroxisomal matrix protein import and lack normal peroxisomes. Instead, they contain peroxisomal membrane remnants, which harbor a very small amount of the major peroxisomal matrix enzymes alcohol oxidase (AO) and dihydroxyacetone synthase (DHAS). The bulk of these proteins is, however, mislocated in the cytosol. Here, we show that in Deltapex14 cells overproduction of the PTS1 receptor, Pex5p, leads to enhanced import of the PTS1 proteins AO and DHAS but not of the PTS2 protein amine oxidase. The import of the PTS1 protein catalase (CAT) was not stimulated by Pex5p overproduction. The difference in import behavior of AO and CAT was not related to their PTS1, since green fluorescent protein fused to the PTS1 of either AO or CAT were both not imported in Deltapex14 cells overproducing Pex5p. When produced in a wild type control strain, both proteins were normally imported into peroxisomes. In Deltapex14 cells overproducing Pex5p, Pex5p had a dual location and was localized in the cytosol and bound to the outer surface of the peroxisomal membrane. Our results indicate that binding of Pex5p to the peroxisomal membrane and import of certain PTS1 proteins can proceed in the absence of Pex14p.
authors.affiliation: Eukaryotic Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, P. O. Box 14, 9750 AA Haren, The Netherlands.
authors.forename: F A
authors.initials: FA
authors.lastname: Salomons
authors.forename: J A
authors.initials: JA
authors.lastname: Kiel
authors.forename: K N
authors.initials: KN
authors.lastname: Faber
authors.forename: M
authors.initials: M
authors.lastname: Veenhuis
authors.forename: I J
authors.initials: IJ
authors.lastname: van der Klei
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D019880:Aldehyde-Ketone Transferases / Q000378:metabolism*; D015153:Blotting, Western; D002352:Carrier Proteins; D010450:Endopeptidases / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology*; D005990:Glycerol / Q000378:metabolism; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D026901:Membrane Transport Proteins; D000432:Methanol / Q000494:pharmacology; D008854:Microscopy, Electron; D008954:Models, Biological; D016296:Mutagenesis; D000074401:Peroxins; D000074435:Peroxisome-Targeting Signal 1 Receptor; D010843:Pichia / Q000166:cytology / Q000235:genetics; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D013395:Sucrose / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D005656:Fungal Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; C499195:PEX14 protein, S cerevisiae; D000074401:Peroxins; D000074435:Peroxisome-Targeting Signal 1 Receptor; D018160:Receptors, Cytoplasmic and Nuclear; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D013395:Sucrose; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D019880:Aldehyde-Ketone Transferases; C031222:formaldehyde transketolase; D010450:Endopeptidases; D005990:Glycerol; D000432:Methanol