The peroxin Pex6p gene is impaired in peroxisomal biogenesis disorders of complementation group 6

Matsumoto, Naomi; Tamura, Shigehiko; Moser, Ann B.; Moser, Hugo W.; Braverman, Nancy; Suzuki, Yasuyuki; Shimozawa, Nobuyuki; Kondo, Naomi; Fujiki, Yukio

doi:10.1007/s100380170078

Cited by 31 publications

(22 citation statements)

References 16 publications

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“…Impairment of Pex1 and Pex6 of the AAA ATPase family (26,40,42,47,52,57) and the recruiter Pex26 of the Pex1-Pex6 complexes (24,25) causes defects in matrix protein import. We investigated whether these peroxins are involved in Pex5 import.…”

Section: Resultsmentioning

confidence: 99%

Shuttling Mechanism of Peroxisome Targeting Signal Type 1 Receptor Pex5: ATP-Independent Import and ATP-Dependent Export

Miyata

Fujiki

2005

Molecular and Cellular Biology

187

218

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Peroxisomal matrix proteins are posttranslationally imported into peroxisomes with the peroxisome-targeting signal 1 receptor, Pex5. The longer isoform of Pex5, Pex5L, also transports Pex7-PTS2 protein complexes. After unloading the cargoes, Pex5 returns to the cytosol. To address molecular mechanisms underlying Pex5 functions, we constructed a cell-free Pex5 translocation system with a postnuclear supernatant fraction from CHO cell lines. In assays using the wild-type CHO-K1 cell fraction, 35 S-labeled Pex5 was specifically imported into and exported from peroxisomes with multiple rounds.35 S-Pex5 import was also evident using peroxisomes isolated from rat liver. ATP was not required for 35 S-Pex5 import but was indispensable for export. 35 S-Pex5 was imported neither to peroxisome remnants from RING peroxin-deficient cell mutants nor to those from pex14 cells lacking a Pex5-docking site. In contrast, 35 S-Pex5 was imported into the peroxisome remnants of PEX1-, PEX6-, and PEX26-defective cell mutants, including those from patients with peroxisome biogenesis disorders, from which, however, 35 S-Pex5 was not exported, thereby indicating that Pex1 and Pex6 of the AAA ATPase family and their recruiter, Pex26, were essential for Pex5 export. Moreover, we analyzed the 35 S-Pex5-associated complexes on peroxisomal membranes by blue-native polyacrylamide gel electrophoresis.35 S-Pex5 was in two distinct, 500-and 800-kDa complexes comprising different sets of peroxins, such as Pex14 and Pex2, implying that Pex5 transited between the subcomplexes. Together, results indicated that Pex5 most likely enters peroxisomes, changes its interacting partners, and then exits using ATP energy.

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Section: Resultsmentioning

confidence: 99%

Shuttling Mechanism of Peroxisome Targeting Signal Type 1 Receptor Pex5: ATP-Independent Import and ATP-Dependent Export

Miyata

Fujiki

2005

Molecular and Cellular Biology

187

218

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“…Peroxisomes in human fibroblasts and CHO cells were visualized by indirect immunofluorescence light microscopy, as described (9). Rabbit antibodies used were antibodies to human catalase (5), peroxisome targeting signal type 1 (PTS1) peptide (22), rat 3-ketoacyl-CoA thiolase (thiolase) (23), and rat Pex14p C-terminal peptide (24).…”

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confidence: 99%

Mutations in the Peroxin Pex26p Responsible for Peroxisome Biogenesis Disorders of Complementation Group 8 Impair Its Stability, Peroxisomal Localization, and Interaction with the Pex1p·Pex6p Complex

Furuki

Tamura

Matsumoto

et al. 2006

Journal of Biological Chemistry

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Peroxisome biogenesis disorders (PBDs) are fatal autosomal recessive diseases and are caused by impaired peroxisome biogenesis. PBDs are genetically heterogeneous and classified into 13 complementation groups (CGs). CG8 is one of the most common groups and has three clinical phenotypes, including Zellweger syndrome (ZS), neonatal adrenoleukodystrophy, and infantile Refsum disease (IRD). We recently isolated PEX26 as the pathogenic gene for PBD of CG8. Pex26p functions in recruiting to peroxisomes the complexes of the AAA ATPase peroxins, Pex1p and Pex6p. In the present work, we identified four distinct mutations in PEX26 from five patients of CG8 PBD including 2 with ZS and 3 with IRD, in addition to 7 mutant alleles in 8 patients in the first report describing the pathogenic PEX26 gene for CG8 PBD. Phenotype-genotype analyses revealed that temperature-sensitive (ts) peroxisome assembly gave rise to a milder IRD in contrast to the non-ts phenotype of the cells from ZS patients. Furthermore, we present several lines of evidence that show that the instability, insufficient binding to Pex1p⅐Pex6p complexes, or mislocalization of patient-derived Pex26p mutants is most likely responsible for the CG8 PBDs.

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“…Peroxisomes were visualized by indirect immunofluorescence light microscopy using rabbit antibodies against human catalase (25), PTS1 peptide (28), and influenza virus hemagglutinin (HA) (29). Antigen-antibody complexes were detected with fluorescein isothiocyanate-labeled goat antibodies against rabbit immunoglobulin G (MP Biomedicals-Cappel, Irvine, CA.…”

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confidence: 99%

“…Plasmids for C-terminal deletion mutants of Pex1p were generated by replacing SpeI-NotI region of full-length PEX1 in pPC86 with SpeI-NotI fragment of the PCR products amplified using a forward primer P1F9 (5Ј-CAAACTTGCCCATACGAC-3Ј) with a reverse P1-1151stopR (5Ј-GCTTGCGGCCGCTTAACA TTGTG-AGCTCAAATC-3Ј) and P1-1217stopR (5Ј-TCCTGCGGC-CGCTTAGGATTCGTCCTCTCCACT-3Ј). To clone the full-length PEX6 in pDB, the HindIII (blunted)-NotI fragment of the FLAG-HsPEX6 in pCMVSPORT (25) was ligated into SalI (blunted) and NotI sites in pDBLeu. For cloning of plasmids for C-terminal deletion mutants, pDB-Pex6p-(1-676) and pDB-Pex6p-(1-882) were generated by replacing BssHII-NotI fragment of pCMVSPORT-HsPEX6-(1-676) and -HsPEX6-(1-882) that had been constructed by replacing the XhoI-NotI fragment of the PCR products amplified using a primer P6F2 (5Ј-GGGCTCGGACCGCGAGTC-3Ј) with P6 -677stopR (5Ј-CTGCGCGGCCGCCTAAGCCAG-GAGAGGAAAGCC-3Ј) and P6 -883stopR (5Ј-TGTGGCGG-CCGCCTAAACGCGTAGCTGGGAGGC-3Ј), respectively.…”

mentioning

confidence: 99%

Dynamic and Functional Assembly of the AAA Peroxins, Pex1p and Pex6p, and Their Membrane Receptor Pex26p

Tamura

Yasutake

Matsumoto

et al. 2006

Journal of Biological Chemistry

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Two AAA peroxins, Pex1p and Pex6p, are encoded by PEX1 and PEX6, the causal genes for peroxisome biogenesis disorders of complementation group 1 (CG1) and CG4, respectively. PEX26 responsible for peroxisome biogenesis disorders of CG8 encodes Pex26p, the recruiter of Pex1p⅐Pex6p complexes to peroxisomes. We herein assigned the binding regions between human Pex1p and Pex6p and elucidated pivotal roles of the AAA cassettes, called D1 and D2 domains, in Pex1p-Pex6p interaction and peroxisome biogenesis. ATP binding in both AAA cassettes but not ATP hydrolysis in D2 of both Pex1p and Pex6p was prerequisite for Pex1p-Pex6p interaction and their peroxisomal localization. The AAA cassettes, D1 and D2, were essential for peroxisome-restoring activity of Pex1p and Pex6p. In HEK293 cells, endogenous Pex1p was partly localized likely as a homo-oligomer in the cytoplasm, while Pex6p and Pex26p were predominantly localized on peroxisomes. Interaction of Pex1p with Pex6p conferred a conformational change and dissociation of the Pex1p oligomer. These results suggested that Pex1p possesses two distinct oligomeric forms, a homo-oligomer in the cytosol and a hetero-oligomer on peroxisome membranes, possibly playing distinct functions in peroxisome biogenesis.

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The peroxin Pex6p gene is impaired in peroxisomal biogenesis disorders of complementation group 6

Cited by 31 publications

References 16 publications

Shuttling Mechanism of Peroxisome Targeting Signal Type 1 Receptor Pex5: ATP-Independent Import and ATP-Dependent Export

Shuttling Mechanism of Peroxisome Targeting Signal Type 1 Receptor Pex5: ATP-Independent Import and ATP-Dependent Export

Mutations in the Peroxin Pex26p Responsible for Peroxisome Biogenesis Disorders of Complementation Group 8 Impair Its Stability, Peroxisomal Localization, and Interaction with the Pex1p·Pex6p Complex

Dynamic and Functional Assembly of the AAA Peroxins, Pex1p and Pex6p, and Their Membrane Receptor Pex26p

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