Alternative Splicing Suggests Extended Function of PEX26 in Peroxisome Biogenesis

• Published in: American journal of human genetics Vol. 76; no. 6; pp. 987 - 1007
• Main Authors: Weller, Sabine, Cajigas, Ivelisse, Morrell, James, Obie, Cassandra, Steel, Gary, Gould, Stephen J., Valle, David
• Format: Journal Article
• Language: English
• Published: Chicago, IL Elsevier Inc 01.06.2005; University of Chicago Press; Cell Press; The American Society of Human Genetics
• Subjects: Alleles; Alternative Splicing; Amino Acid Sequence - genetics; Biological and medical sciences; Cell Line; Chromosome Mapping; Cytosol - metabolism; Disease; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Exons; Fibroblasts - metabolism; Gene Frequency; General aspects. Genetic counseling; Genetics; Genotype & phenotype; Humans; Medical genetics; Medical sciences; Membrane Proteins - chemistry; Membrane Proteins - genetics; Membrane Proteins - metabolism; Metabolism; Mitochondria - metabolism; Models, Genetic; Molecular Sequence Data; Mutation, Missense; Pedigree; Peroxisomal Disorders - genetics; Peroxisomal Disorders - metabolism; Peroxisomal Targeting Signal 2 Receptor; Peroxisome-Targeting Signal 1 Receptor; Peroxisomes - chemistry; Peroxisomes - metabolism; Protein Isoforms - chemistry; Protein Isoforms - genetics; Protein Isoforms - metabolism; Protein Structure, Tertiary; Proteins; Receptors, Cytoplasmic and Nuclear - genetics; Receptors, Cytoplasmic and Nuclear - metabolism; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; Sequence Homology, Amino Acid; Skin - cytology; Human; Peroxisome; Alternative splicing; Genetics
• ISSN: 0002-9297; 1537-6605
• DOI: 10.1086/430637

Matsumoto and colleagues recently identified PEX26 as the gene responsible for complementation group 8 of the peroxisome biogenesis disorders and showed that it encodes an integral peroxisomal membrane protein with a single C-terminal transmembrane domain and a cytosolic N-terminus that interacts with the PEX1/PEX6 heterodimer through direct binding to the latter. They proposed that PEX26 functions as the peroxisomal docking factor for the PEX1/PEX6 heterodimer. Here, we identify new PEX26 disease alleles, localize the PEX6-binding domain to the N-terminal half of the protein (aa 29–174), and show that, at the cellular level, PEX26 deficiency impairs peroxisomal import of both PTS1- and PTS2-targeted matrix proteins. Also, we find that PEX26 undergoes alternative splicing to produce several splice forms—including one, PEX26-Δex5, that maintains frame and encodes an isoform lacking the transmembrane domain of full-length PEX26 (PEX26-FL). Despite its cytosolic location, PEX26-Δex5 rescues peroxisome biogenesis in PEX26-deficient cells as efficiently as does PEX26-FL. To test our observation that a peroxisomal location is not required for PEX26 function, we made a chimeric protein (PEX26-Mito) with PEX26 as its N-terminus and the targeting segment of a mitochondrial outer membrane protein (OMP25) at its C-terminus. We found PEX26-Mito localized to the mitochondria and directed all detectable PEX6 and a fraction of PEX1 to this extraperoxisomal location; yet PEX26-Mito retains the full ability to rescue peroxisome biogenesis in PEX26-deficient cells. On the basis of these observations, we suggest that a peroxisomal localization of PEX26 and PEX6 is not required for their function and that the interaction of PEX6 with PEX1 is dynamic. This model predicts that, once activated in an extraperoxisomal location, PEX1 moves to the peroxisome and completes the function of the PEX1/6 heterodimer.