Structural Insights into Cargo Recognition by the Yeast PTS1 Receptor

Hagen, Stefanie; Drepper, Friedel; Fischer, Sabrina; Fodor, Krisztián; Passon, Daniel M.; Platta, Harald W.; Zenn, Michael; Schliebs, Wolfgang; Girzalsky, Wolfgang; Wilmanns, Matthias; Warscheid, Bettina; Erdmann, Ralf

doi:10.1074/jbc.m115.657973

Cited by 28 publications

(29 citation statements)

References 68 publications

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“…In contrast, efficient binding was observed when GST-Pex5p was bound to the agarose ( Fig. 4A), which is in line with our previous reports of a direct Pex5p/Pcs60p interaction (37). Pcs60p was also coeluted when GST-Ub (⌬1-6) Pex5p was bound to the column.…”

Section: Ubiquitin-dependent Receptor Recognition Mediated By Pex1psupporting

confidence: 92%

“…crucial for peroxisomal targeting (35). Moreover, a direct binding to the PTS1 receptor was previously reported (37). To investigate the influence of the linear fusion of ubiquitin to the N terminus of Pex5p on receptor cargo recognition, we tested whether presence of the ubiquitin moiety alters the in vitro binding of Ub-Pex5p to Pcs60p.…”

Section: Ubiquitin-dependent Receptor Recognition Mediated By Pex1pmentioning

confidence: 99%

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Receptor recognition by the peroxisomal AAA complex depends on the presence of the ubiquitin moiety and is mediated by Pex1p

Schwerter

Grimm

Girzalsky

et al. 2018

Journal of Biological Chemistry

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The receptor cycle of type I peroxisomal matrix protein import is completed by ubiquitination of the membrane-bound peroxisome biogenesis factor 5 (Pex5p) and its subsequent export back to the cytosol. The receptor export is the only ATP-dependent step of the whole process and is facilitated by two members of the AAA family of proteins (ATPases associated with various cellular activities), namely Pex1p and Pex6p. To gain further insight into substrate recognition by the AAA complex, we generated an N-terminally linked ubiquitin-Pex5p fusion protein. This fusion protein displayed biological activity because it is able to functionally complement a PEX5-deletion in assays revealed its interaction at WT level with the native cargo protein Pcs60p and Pex14p, a constituent of the receptor docking complex. We also demonstrate deubiquitination by the deubiquitinating enzyme Ubp15p. pulldown assays and cross-linking studies demonstrate that Pex5p recognition by the AAA complex depends on the presence of the ubiquitin moiety and is mediated by Pex1p.

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Section: Ubiquitin-dependent Receptor Recognition Mediated By Pex1psupporting

confidence: 92%

Section: Ubiquitin-dependent Receptor Recognition Mediated By Pex1pmentioning

confidence: 99%

Receptor recognition by the peroxisomal AAA complex depends on the presence of the ubiquitin moiety and is mediated by Pex1p

Schwerter

Grimm

Girzalsky

et al. 2018

Journal of Biological Chemistry

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“…This construct still contains the TPR domains, which, in Pex5p, are known to form a PTS1-binding domain (Gatto et al, 2000). Interestingly, another peroxisomal protein, Pcs60p, which also harbors the C-terminal targeting sequence SKL and which is known to interact with Pex5p (Hagen et al, 2015), did not interact with Pex9p. In addition, no interaction between Fox1p, a Pex5p-dependent non-PTS cargo protein, and Pex9p could be detected.…”

Section: Pex9p Interacts With Pex14p and Peroxisomal Pts1-containing mentioning

confidence: 99%

Pex9p is a new yeast peroxisomal import receptor for PTS1-containing proteins

Effelsberg

Cruz-Zaragoza

Schliebs

et al. 2016

Journal of Cell Science

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Peroxisomal proteins carrying a type 1 peroxisomal targeting signal (PTS1) are recognized by the well-conserved cycling import receptor Pex5p. The yeast YMR018W gene encodes a Pex5p paralog and newly identified peroxin that is involved in peroxisomal import of a subset of matrix proteins. The new peroxin was designated Pex9p, and it interacts with the docking protein Pex14p and a subclass of PTS1-containing peroxisomal matrix enzymes. Unlike Pex5p, Pex9p is not expressed in glucose-or ethanol-grown cells, but it is strongly induced by oleate. Under these conditions, Pex9p acts as a cytosolic and membrane-bound peroxisome import receptor for both malate synthase isoenzymes, Mls1p and Mls2p. The inducible Pex9p-dependent import pathway provides a mechanism for the oleate-inducible peroxisomal targeting of malate synthases. The existence of two distinct PTS1 receptors, in addition to two PTS2-dependent import routes, contributes to the adaptive metabolic capacity of peroxisomes in response to environmental changes and underlines the role of peroxisomes as multi-purpose organelles. The identification of different import routes into peroxisomes contributes to the molecular understanding of how regulated protein targeting can alter the function of organelles according to cellular needs.

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“…Interactions of PEX5 with other peroxins can affect PTS1 binding, as shown for PEX13 influencing the import of catalase (ending in a weak PTS1:KANL) (8). For some PTS1 proteins, secondary interactions with PEX5 have been described (9). More recently, redox reactions involving a cysteine group in the N terminus of the protein were shown to modulate the affinity of PEX5 for its targets (10,11).…”

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confidence: 99%

Allostery between two binding sites in the ion channel subunit TRIP8b confers binding specificity to HCN channels

Lyman

Han

Heuermann

et al. 2017

Journal of Biological Chemistry

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Tetratricopeptide repeat (TPR) domains are ubiquitous structural motifs that mediate protein-protein interactions. For example, the TPR domains in the peroxisomal import receptor PEX5 enable binding to a range of type 1 peroxisomal targeting signal motifs. A homolog of PEX5, tetratricopeptide repeat-containing Rab8b-interacting protein (TRIP8b), binds to and functions as an auxiliary subunit of hyperpolarization-activated cyclic nucleotide (HCN)-gated channels. Given the similarity between TRIP8b and PEX5, this difference in function raises the question of what mechanism accounts for their binding specificity. In this report, we found that the cyclic nucleotide-binding domain and the C terminus of the HCN channel are critical for conferring specificity to TRIP8b binding. We show that TRIP8b binds the HCN cyclic nucleotide-binding domain through a 37-residue domain and the HCN C terminus through the TPR domains. Using a combination of fluorescence polarization- and co-immunoprecipitation-based assays, we establish that binding at either site increases affinity at the other. Thus, allosteric coupling of the TRIP8b TPR domains both promotes binding to HCN channels and limits binding to type 1 peroxisomal targeting signal substrates. These results raise the possibility that other TPR domains may be similarly influenced by allosteric mechanisms as a general feature of protein-protein interactions.

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Structural Insights into Cargo Recognition by the Yeast PTS1 Receptor

Cited by 28 publications

References 68 publications

Receptor recognition by the peroxisomal AAA complex depends on the presence of the ubiquitin moiety and is mediated by Pex1p

Receptor recognition by the peroxisomal AAA complex depends on the presence of the ubiquitin moiety and is mediated by Pex1p

Pex9p is a new yeast peroxisomal import receptor for PTS1-containing proteins

Allostery between two binding sites in the ion channel subunit TRIP8b confers binding specificity to HCN channels

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