title: Different functions of the C3HC4 zinc RING finger peroxins PEX10, PEX2, and PEX12 in peroxisome formation and matrix protein import.
abstract: The integral peroxisomal membrane proteins PEX10, PEX2, and PEX12 contain a zinc RING finger close to the C terminus. Loss of function of these peroxins causes embryo lethality at the heart stage in Arabidopsis. Preventing the coordination of Zn(2+) ions by amino acid substitutions in PEX10, PEX2, and PEX12 and overexpressing the resulting conditional sublethal mutations in WT uncovered additional functions of PEX10. Plants overexpressing DeltaZn-mutant PEX10 display deformed peroxisomal shapes causing diminished contact with chloroplasts and possibly with mitochondria. These changes correlated with impaired metabolite transfer and, at high CO(2), recoverable defective photorespiration plus dwarfish phenotype. The N-terminal PEX10 domain is critical for peroxisome biogenesis and plant development. A point mutation in the highly conserved TLGEEY motif results in vermiform peroxisome shape without impairing organelle contact. Addition of an N-terminal T7 tag to WT PEX0 resulted in partially recoverable reduced growth and defective inflorescences persisting under high CO(2). In contrast, plants overexpressing PEX2-DeltaZn-T7 grow like WT in normal atmosphere, contain normal-shaped peroxisomes, but display impaired peroxisomal matrix protein import. PEX12-DeltaZn-T7 mutants exhibit unimpaired import of matrix protein and normal-shaped peroxisomes when grown in normal atmosphere. During seed germination, glyoxysomes form a reticulum around the lipid bodies for mobilization of storage oil. The formation of this glyoxysomal reticulum seemed to be impaired in PEX10-DeltaZn but not in PEX2-DeltaZn-T7 or PEX12-DeltaZn-T7 plants. Both cytosolic PEX10 domains seem essential for peroxisome structure but differ in metabolic function, suggesting a role for this plant peroxin in addition to the import of matrix protein via ubiquitination of PEX5.
authors.affiliation: LS Botanik and LS Systembiologie der Pflanzen, Technische Universität München, D-85350 Freising, Germany.
authors.forename: Jakob
authors.initials: J
authors.lastname: Prestele
authors.forename: Georg
authors.initials: G
authors.lastname: Hierl
authors.forename: Christian
authors.initials: C
authors.lastname: Scherling
authors.forename: Stefan
authors.initials: S
authors.lastname: Hetkamp
authors.forename: Claus
authors.initials: C
authors.lastname: Schwechheimer
authors.forename: Erika
authors.initials: E
authors.lastname: Isono
authors.forename: Wolfram
authors.initials: W
authors.lastname: Weckwerth
authors.forename: Gerhard
authors.initials: G
authors.lastname: Wanner
authors.forename: Christine
authors.initials: C
authors.lastname: Gietl
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D001692:Biological Transport; D002245:Carbon Dioxide / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D020676:Glyoxysomes / Q000378:metabolism / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D055432:Metabolomics / Q000379:methods; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D000074401:Peroxins; D000074435:Peroxisome-Targeting Signal 1 Receptor; D020675:Peroxisomes / Q000378:metabolism* / Q000648:ultrastructure; D010788:Photosynthesis; D030821:Plants, Genetically Modified; D054829:RING Finger Domains / Q000235:genetics; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D016335:Zinc Fingers / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; D016326:Extracellular Matrix Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; C477605:PEX10 protein, Arabidopsis; D000074401:Peroxins; D000074435:Peroxisome-Targeting Signal 1 Receptor; D018160:Receptors, Cytoplasmic and Nuclear; C462525:TED3 protein, Arabidopsis; C507147:peroxin 12 protein, Arabidopsis; D002245:Carbon Dioxide; D049452:Green Fluorescent Proteins