title: Pnc1 piggy-back import into peroxisomes relies on Gpd1 homodimerisation.
abstract: Peroxisomes are eukaryotic organelles that posttranslationally import proteins via one of two conserved peroxisomal targeting signal (PTS1 or 2) mediated pathways. Oligomeric proteins can be imported via these pathways but evidence is accumulating that at least some PTS1-containing monomers enter peroxisomes before they assemble into oligomers. Some proteins lacking a PTS are imported by piggy-backing onto PTS-containing proteins. One of these proteins is the nicotinamidase Pnc1, that is co-imported with the PTS2-containing enzyme Glycerol-3-phosphate dehydrogenase 1, Gpd1. Here we show that Pnc1 co-import requires Gpd1 to form homodimers. A mutation that interferes with Gpd1 homodimerisation does not prevent Gpd1 import but prevents Pnc1 co-import. A suppressor mutation that restores Gpd1 homodimerisation also restores Pnc1 co-import. In line with this, Pnc1 interacts with Gpd1 in vivo only when Gpd1 can form dimers. Redirection of Gpd1 from the PTS2 import pathway to the PTS1 import pathway supports Gpd1 monomer import but not Gpd1 homodimer import and Pnc1 co-import. Our results support a model whereby Gpd1 may be imported as a monomer or a dimer but only the Gpd1 dimer facilitates co-transport of Pnc1 into peroxisomes.
authors.affiliation: Department of Molecular Biology and Biotechnology University of Sheffield Firth Court, Western Bank Sheffield S10 2TN United Kingdom.
authors.forename: Nadal A Al
authors.initials: NA
authors.lastname: Saryi
authors.affiliation: Department of Molecular Biology and Biotechnology University of Sheffield Firth Court, Western Bank Sheffield S10 2TN United Kingdom.
authors.forename: John D
authors.initials: JD
authors.lastname: Hutchinson
authors.affiliation: Department of Molecular Biology and Biotechnology University of Sheffield Firth Court, Western Bank Sheffield S10 2TN United Kingdom.
authors.forename: Murtakab Y
authors.initials: MY
authors.lastname: Al-Hejjaj
authors.affiliation: Department of Molecular Biology and Biotechnology University of Sheffield Firth Court, Western Bank Sheffield S10 2TN United Kingdom.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Sedelnikova
authors.affiliation: Department of Molecular Biology and Biotechnology University of Sheffield Firth Court, Western Bank Sheffield S10 2TN United Kingdom.
authors.forename: Patrick
authors.initials: P
authors.lastname: Baker
authors.affiliation: Department of Molecular Biology and Biotechnology University of Sheffield Firth Court, Western Bank Sheffield S10 2TN United Kingdom.
authors.forename: Ewald H
authors.initials: EH
authors.lastname: Hettema
mesh_terms: D015870:Gene Expression; D017930:Genes, Reporter; D005993:Glycerolphosphate Dehydrogenase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D033681:Mitochondrial Membrane Transport Proteins; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D009154:Mutation; D033704:Nucleotide Transport Proteins / Q000235:genetics / Q000378:metabolism*; D020675:Peroxisomes / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011487:Protein Conformation; D055503:Protein Multimerization; D021381:Protein Transport; D015398:Signal Transduction
chemical_list: D033681:Mitochondrial Membrane Transport Proteins; D024101:Mitochondrial Proteins; D033704:Nucleotide Transport Proteins; C483480:SLC25A33 protein, human; D005993:Glycerolphosphate Dehydrogenase