PER3, a Gene Required for Peroxisome Biogenesis inPichia pastoris, Encodes a Peroxisomal Membrane Protein Involved in Protein Import

We identified a Saccharomyces cerevisiae peroxisomal membrane protein, Pex13p, that is essential for protein import. A point mutation in the COOH-terminal Src homology 3 (SH3) domain of Pex13p inactivated the protein but did not affect its membrane targeting. A two-hybrid screen with the SH3 domain of Pex13p identified Pex5p, a receptor for proteins with a type I peroxisomal targeting signal (PTS1), as its ligand. Pex13p SH3 interacted specifically with Pex5p in vitro. We determined, furthermore, that Pex5p was mainly present in the cytosol and only a small fraction was associated with peroxisomes. We therefore propose that Pex13p is a component of the peroxisomal protein import machinery onto which the mobile Pex5p receptor docks for the delivery of the selected PTS1 protein.

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Peroxisomal Membrane Protein Import Does Not Require Pex17p

Journal of Biological Chemistry ◽

10.1074/jbc.m111728200 ◽

2002 ◽

Vol 277 (19) ◽

pp. 16498-16504 ◽

Cited By ~ 15

Author(s):

Courtney C. Harper ◽

Sarah T. South ◽

J. Michael McCaffery ◽

Stephen J. Gould

Keyword(s):

Membrane Protein ◽

Protein Import ◽

Peroxisomal Membrane ◽

Peroxisomal Membrane Protein

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The importomer is a peroxisomal membrane protein translocase

10.1101/2020.05.01.072660 ◽

2020 ◽

Author(s):

James S Martenson ◽

Hanson Tam ◽

Alexander J McQuown ◽

Dvir Reif ◽

Jing Zhou ◽

...

Keyword(s):

Membrane Protein ◽

Matrix Protein ◽

Protein Import ◽

Protein Targeting ◽

Fundamental Principle ◽

Specific Protein ◽

En Bloc ◽

Peroxisomal Membrane ◽

Peroxisomal Membrane Protein ◽

Protein Biogenesis

ABSTRACTThree sites of membrane protein biogenesis (the endoplasmic reticulum, mitochondria and chloroplasts) receive unfolded substrates from organelle-specific protein targeting factors, then integrate them using separate translocation channels. Peroxisomes also receive membrane proteins from known targeting factors, but whether a separate translocase is needed for integration remains unknown. Here, using a novel genetic screening strategy, we reveal that the importomer–known for matrix protein import–integrates the peroxisomal membrane protein Pex14. In importomer mutants, Pex14 is arrested in a pre-integrated state on the peroxisome surface. To undergo integration, a Pex14 translocation signal binds the importomer’s substrate receptor Pex5 at a distinct site from matrix proteins. En bloc translocation of an engineered protein complex with Pex14’s luminal region argues that integration occurs without substrate unfolding. Our work shows that the handling of membrane protein targeting and integration by discrete machineries is a fundamental principle shared by diverse membrane protein biogenesis pathways.

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Pex12 Interacts with Pex5 and Pex10 and Acts Downstream of Receptor Docking in Peroxisomal Matrix Protein Import

The Journal of Cell Biology ◽

10.1083/jcb.147.4.761 ◽

1999 ◽

Vol 147 (4) ◽

pp. 761-774 ◽

Cited By ~ 123

Author(s):

Chia-Che Chang ◽

Daniel S. Warren ◽

Katherine A. Sacksteder ◽

Stephen J. Gould

Keyword(s):

Membrane Protein ◽

Matrix Protein ◽

Protein Import ◽

Zellweger Syndrome ◽

Binding Domain ◽

Ring Domain ◽

Zinc Binding ◽

Peroxisomal Membrane ◽

Peroxisomal Membrane Protein ◽

Zinc Binding Domain

Peroxisomal matrix protein import requires PEX12, an integral peroxisomal membrane protein with a zinc ring domain at its carboxy terminus. Mutations in human PEX12 result in Zellweger syndrome, a lethal neurological disorder, and implicate the zinc ring domain in PEX12 function. Using two-hybrid studies, blot overlay assays, and coimmunoprecipitation experiments, we observed that the zinc-binding domain of PEX12 binds both PEX5, the PTS1 receptor, and PEX10, another integral peroxisomal membrane protein required for peroxisomal matrix protein import. Furthermore, we identified a patient with a missense mutation in the PEX12 zinc-binding domain, S320F, and observed that this mutation reduces the binding of PEX12 to PEX5 and PEX10. Overexpression of either PEX5 or PEX10 can suppress this PEX12 mutation, providing genetic evidence that these interactions are biologically relevant. PEX5 is a predominantly cytoplasmic protein and previous PEX5-binding proteins have been implicated in docking PEX5 to the peroxisome surface. However, we find that loss of PEX12 or PEX10 does not reduce the association of PEX5 with peroxisomes, demonstrating that these peroxins are not required for receptor docking. These and other results lead us to propose that PEX12 and PEX10 play direct roles in peroxisomal matrix protein import downstream of the receptor docking event.

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Isolation and Characterization of Pas2p, a Peroxisomal Membrane Protein Essential for Peroxisome Biogenesis in the Methylotrophic YeastPichia pastoris

Journal of Biological Chemistry ◽

10.1074/jbc.271.31.18973 ◽

1996 ◽

Vol 271 (31) ◽

pp. 18973-18980 ◽

Cited By ~ 93

Author(s):

Erik A. C. Wiemer ◽

Georg H. Lüers ◽

Klaas Nico Faber ◽

Thibaut Wenzel ◽

Marten Veenhuis ◽

...

Keyword(s):

Membrane Protein ◽

Peroxisome Biogenesis ◽

Peroxisomal Membrane ◽

Peroxisomal Membrane Protein ◽

Isolation And Characterization

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The 70 kDa peroxisomal membrane protein: an ATP-binding cassette transporter protein involved in peroxisome biogenesis

Seminars in Cell Biology ◽

10.1006/scel.1993.1006 ◽

1993 ◽

Vol 4 (1) ◽

pp. 45-52 ◽

Cited By ~ 18

Author(s):

Jutta Gärtner ◽

David Valle

Keyword(s):

Membrane Protein ◽

Atp Binding ◽

Peroxisome Biogenesis ◽

Peroxisomal Membrane ◽

Peroxisomal Membrane Protein ◽

Transporter Protein ◽

Atp Binding Cassette Transporter ◽

Atp Binding Cassette

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Peroxisome Synthesis in the Absence of Preexisting Peroxisomes

The Journal of Cell Biology ◽

10.1083/jcb.144.2.255 ◽

1999 ◽

Vol 144 (2) ◽

pp. 255-266 ◽

Cited By ~ 173

Author(s):

Sarah T. South ◽

Stephen J. Gould

Keyword(s):

Membrane Proteins ◽

Membrane Protein ◽

Matrix Protein ◽

Protein Import ◽

Zellweger Syndrome ◽

Matrix Proteins ◽

Peroxisomal Membrane ◽

Peroxisomal Membrane Protein ◽

The Matrix ◽

Inactivating Mutations

Zellweger syndrome and related diseases are caused by defective import of peroxisomal matrix proteins. In all previously reported Zellweger syndrome cell lines the defect could be assigned to the matrix protein import pathway since peroxisome membranes were present, and import of integral peroxisomal membrane proteins was normal. However, we report here a Zellweger syndrome patient (PBD061) with an unusual cellular phenotype, an inability to import peroxisomal membrane proteins. We also identified human PEX16, a novel integral peroxisomal membrane protein, and found that PBD061 had inactivating mutations in the PEX16 gene. Previous studies have suggested that peroxisomes arise from preexisting peroxisomes but we find that expression of PEX16 restores the formation of new peroxisomes in PBD061 cells. Peroxisome synthesis and peroxisomal membrane protein import could be detected within 2–3 h of PEX16 injection and was followed by matrix protein import. These results demonstrate that peroxisomes do not necessarily arise from division of preexisting peroxisomes. We propose that peroxisomes may form by either of two pathways: one that involves PEX11-mediated division of preexisting peroxisomes, and another that involves PEX16-mediated formation of peroxisomes in the absence of preexisting peroxisomes.

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Pex24 and Pex32 tether peroxisomes to the ER for organelle biogenesis, positioning and segregation

10.1101/2020.03.05.977884 ◽

2020 ◽

Author(s):

Fei Wu ◽

Rinse de Boer ◽

Arjen M. Krikken ◽

Arman Akşit ◽

Nicola Bordin ◽

...

Keyword(s):

Membrane Protein ◽

Hansenula Polymorpha ◽

Peroxisome Biogenesis ◽

Organelle Biogenesis ◽

Peroxisomal Membrane ◽

Peroxisomal Protein ◽

Peroxisomal Membrane Protein ◽

Contact Formation ◽

And Function

AbstractWe analyzed all four Pex23 family proteins of the yeast Hansenula polymorpha, which localize to the ER. Of these Pex24 and Pex32, but not Pex23 and Pex29, accumulate at peroxisome-ER contacts, where they are important for normal peroxisome biogenesis and proliferation and contribute to organelle positioning and segregation.Upon deletion of PEX24 and PEX32 - and to a lesser extent of PEX23 and PEX29 - peroxisome-ER contacts are disrupted, concomitant with peroxisomal defects. These defects are suppressed upon introduction of an artificial peroxisome-ER tether.Accumulation of Pex32 at peroxisomes-ER contacts is lost in the absence of the peroxisomal membrane protein Pex11. At the same time peroxisome-ER contacts are disrupted, indicating that Pex11 contributes to Pex32-dependent peroxisome-ER contact formation.Summarizing, our data indicate that H. polymorpha Pex24 and Pex32 are tethers at peroxisome-ER contacts that are important for normal peroxisome biogenesis and dynamics.SummaryTwo Hansenula polymorpha ER proteins, Pex24 and Pex32, are tethers at peroxisome-ER contacts and function together with the peroxisomal protein Pex11. Their absence disturbs these contacts leading to multiple peroxisomal defects, which can be restored by an artificial tether.

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Identification of Pex13p a peroxisomal membrane receptor for the PTS1 recognition factor.

The Journal of Cell Biology ◽

10.1083/jcb.135.1.111 ◽

1996 ◽

Vol 135 (1) ◽

pp. 111-121 ◽

Cited By ~ 167

Author(s):

R Erdmann ◽

G Blobel

Keyword(s):

Membrane Protein ◽

Protein Import ◽

Signal Sequence ◽

Membrane Receptor ◽

Matrix Proteins ◽

Signal Recognition ◽

Peroxisomal Membrane ◽

Peroxisomal Membrane Protein ◽

Src Homology 3 ◽

Src Homology

We have identified an S. cerevisiae integral peroxisomal membrane protein of M of 42,705 (Pex13p) that is a component of the peroxisomal protein import apparatus. Pex13p's most striking feature is an src homology 3 (SH3) domain that interacts directly with yeast Pex5p (former Pas10p), the recognition factor for the COOH-terminal tripeptide signal sequence (PTS1), but not with Pex7p (former Pas7p), the recognition factor for the NH2-terminal nonapeptide signal (PTS2) of peroxisomal matrix proteins. Hence, Pex13p serves as peroxisomal membrane receptor for at least one of the two peroxisomal signal recognition factors. Cells deficient in Pex13p are unable to import peroxisomal matrix proteins containing PTS1 and, surprisingly, also those containing PTS2. Pex13p deficient cells retain membranes containing the peroxisomal membrane protein Pex11p (former Pmp27p), consistent with the existence of independent pathways for the integration of peroxisomal membrane proteins and for the translocation of peroxisomal matrix proteins.

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