The AAA peroxins, Pex1p and Pex6p, are components of the peroxisomal protein import machinery required for the relocation of the import receptor Pex5p from the peroxisomal membrane to the cytosol. We demonstrate that Pex1p and Pex6p form a stable complex in the cytosol, which associates at the peroxisomal membrane with their membrane anchor Pex15p and the peroxisomal importomer. The interconnection of Pex15p with the components of the importomer was independent of Pex1p and Pex6p, indicating that Pex15p is an incorporated component of the assembly. Further evidence suggests that the AAA peroxins shuttle between cytosol and peroxisome with proper binding of the Pex15p-AAA complex to the importomer and release of the AAA peroxins from the peroxisomal membrane depending on an operative peroxisomal protein import mechanism. Pex4p-deficient cells exhibit a wild-type-like assembly of the importomer, which differs in that it is associated with increased amounts of Pex1p and Pex6p, in agreement with a function for Pex4p in the release of AAA peroxins from the peroxisomal membrane.

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http://dx.doi.org/10.1111/j.1742-4658.2006.05388.xDOI Listing

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Article Synopsis
  • - Peroxisomes are essential organelles for lipid metabolism and maintaining redox balance in cells, primarily importing matrix proteins via a process that requires ATP but not right at the start.
  • - The ATP-dependent phase occurs later in the import process, involving a complex interplay between peroxisomal targeting signal (PTS) receptors and the peroxisomal export complex.
  • - Key steps include the monoubiquitination of PTS receptors, facilitating their retrieval from the membrane, and upon deubiquitination, the receptors can help import more proteins into the peroxisome.
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The AAA ATPases PEX1•PEX6 extract PEX5, the peroxisomal protein shuttling receptor, from the peroxisomal membrane so that a new protein transport cycle can start. Extraction requires ubiquitination of PEX5 at residue 11 and involves a threading mechanism, but how exactly this occurs is unclear. We used a cell-free in vitro system and a variety of engineered PEX5 and ubiquitin molecules to challenge the extraction machinery.

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