Dataset Information

1 Structure of the peroxisomal retro-translocon formed by a heterotrimeric ubiquitin ligase 2 complex

ABSTRACT: Peroxisomes are ubiquitous organelles that house various metabolic reactions and are essential for human health. Luminal peroxisomal proteins are imported from the cytosol by mobile receptors, which then recycle back to the cytosol by a poorly understood process. Recycling requires receptor modification by a membrane-embedded ubiquitin ligase complex comprised of three RING finger (RF) domain-containing proteins (Pex2, Pex10, and Pex12). Here, we report a cryo-electron microscopy (cryo-EM) structure of the ligase complex, which together with biochemical and in vivo experiments reveals its function as a retro-translocon for peroxisomal import receptors. Each subunit of the complex contributes five transmembrane segments that co-assemble into an open channel. The three RFs form a cytosolic tower, with RF2 positioned above the channel pore. We propose that the N-terminus of a recycling receptor is inserted from the peroxisomal lumen into the pore and modified by RF2 to enable extraction into the cytosol. If recycling is compromised, receptors are polyubiquitinated by the concerted action of RF10 and RF12 and degraded. This polyubiquitination pathway also maintains the homeostasis of other peroxisomal import factors. Our results clarify a crucial step during peroxisomal protein import and reveal why mutations in the ligase complex cause human disease.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Joao Paulo

LAB HEAD: Tom Rapoport

PROVIDER: PXD031792 | Pride | 2022-07-20

REPOSITORIES: Pride

ACCESS DATA

Publications

A peroxisomal ubiquitin ligase complex forms a retrotranslocation channel.

Feng Peiqiang P Wu Xudong X Erramilli Satchal K SK Paulo Joao A JA Knejski Pawel P Gygi Steven P SP Kossiakoff Anthony A AA Rapoport Tom A TA

Nature 20220629 7918

Peroxisomes are ubiquitous organelles that house various metabolic reactions and are essential for human health1-4. Luminal peroxisomal proteins are imported from the cytosol by mobile receptors, which then recycle back to the cytosol by a poorly understood process1-4. Recycling requires receptor modification by a membrane-embedded ubiquitin ligase complex comprising three RING finger domain-containing proteins (Pex2, Pex10 and Pex12)5,6. Here we report a cryo-el ...[more]

PMID: 35768507

Similar Datasets

Project description:Peroxisome biogenesis diseases (PBDs) are characterized by global defects in peroxisomal function and can result in severe brain, liver, kidney, and bone malfunctions. PBDs are due to mutations in peroxisome biogenesis factors (PEX genes) that are responsible for peroxisome assembly and function. Increasing evidence suggests that peroxisome import functions decline during aging. However, the transcriptome profiling of peroxisome import defects and how they affect disease development are still lacking. PEX5 encodes the cytoplasmic receptors for peroxisome-targeting signal types 1. We generate knock-in human HEK293 cells mutant using CRISPR to transiently express PEX5 cysteine 11 to alanine mutant (PEX5C11A), which blocks PEX5 recycling and exerts dominant negative effect on PEX5 mediated peroxisome import. To identify conserved responses, we perform transcriptomic analysis on Drosophila oenocyte-specific Pex1, Pex12 and Pex5 knockdowns and on human cells with impaired peroxisome import (through expressing PEX5C11A and applying PEX5 siRNA respectively). PEX5C11A induction triggers vast transcriptomic changes, including decreased oxidative phosphorylation, increased MAPK signaling and HIPPO signaling. PEX5 siRNA specifically decreases spliceosome activity and increases cholesterol metabolism. Using gene set enrichment analysis (GSEA), we identify protein processing in endoplasmic reticulum pathway, specifically ER-associated protein degradation (ERAD) pathway is induced in all PEX knockdowns in Drosophila. Peroxisome dysfunction elevates eIF2α phosphorylation in both Drosophila and human cell culture independent of XBP1 activation, suggesting increased integrative stress response (ISR). Moreover, peroxisome stress decreases ribosome biogenesis genes and impairs ribosome biogenesis in flies and human cells. Specifically, peroxisome stress impairs the 5'-ETS cleavage activity during the ribosome biogenesis and dampens 40S small ribosomal export in both flies and human. Our results suggest that reduced ribosome biogenesis and elevated ISR could be conserved cellular response to peroxisome import stress.

Dataset Information

1 Structure of the peroxisomal retro-translocon formed by a heterotrimeric ubiquitin ligase 2 complex

Publications

A peroxisomal ubiquitin ligase complex forms a retrotranslocation channel.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure