Peroxisomes are organelles with vital functions in metabolism and their dysfunction is associated with human diseases. To fulfill their multiple roles, peroxisomes import nuclear-encoded matrix proteins, most carrying a peroxisomal targeting signal (PTS) 1. The receptor Pex5p recruits PTS1-proteins for import into peroxisomes; whether and how this process is posttranslationally regulated is unknown. Here, we identify 22 phosphorylation sites of Pex5p. Yeast cells expressing phospho-mimicking Pex5p-S507/523D (Pex5p(2D)) show decreased import of GFP with a PTS1. We show that the binding affinity between a PTS1-protein and Pex5p(2D) is reduced. An in vivo analysis of the effect of the phospho-mimicking mutant on PTS1-proteins revealed that import of most, but not all, cargos is affected. The physiological effect of the phosphomimetic mutations correlates with the binding affinity of the corresponding extended PTS1-sequences. Thus, we report a novel Pex5p phosphorylation-dependent mechanism for regulating PTS1-protein import into peroxisomes. In a broader view, this suggests that posttranslational modifications can function in fine-tuning the peroxisomal protein composition and, thus, cellular metabolism.
%0 Journal Article
%1 fischerPhosphorylationReceptorProtein2023a
%A Fischer, Sven
%A Bürgi, Jérôme
%A Gabay-Maskit, Shiran
%A Maier, Renate
%A Mastalski, Thomas
%A Yifrach, Eden
%A Obarska-Kosinska, Agnieszka
%A Rudowitz, Markus
%A Erdmann, Ralf
%A Platta, Harald W.
%A Wilmanns, Matthias
%A Schuldiner, Maya
%A Zalckvar, Einat
%A Oeljeklaus, Silke
%A Drepper, Friedel
%A Warscheid, Bettina
%C Germany
%D 2023
%J Biological chemistry
%K *Peroxisomes/metabolism,*Receptors 1 Cytoplasmic Nuclear/metabolism,Carrier Proteins/metabolism,high-content Receptor/metabolism,Pex5p Signal TPR Transport,Saccharomyces and cerevisiae/metabolism,to_read domain,Phosphorylation,posttranslational localization,Protein modification,protein screen,Humans,mass spectrometry,Peroxisome-Targeting
%N 2-3
%P 135--155
%R 10.1515/hsz-2022-0168
%T Phosphorylation of the Receptor Protein Pex5p Modulates Import of Proteins into Peroxisomes.
%V 404
%X Peroxisomes are organelles with vital functions in metabolism and their dysfunction is associated with human diseases. To fulfill their multiple roles, peroxisomes import nuclear-encoded matrix proteins, most carrying a peroxisomal targeting signal (PTS) 1. The receptor Pex5p recruits PTS1-proteins for import into peroxisomes; whether and how this process is posttranslationally regulated is unknown. Here, we identify 22 phosphorylation sites of Pex5p. Yeast cells expressing phospho-mimicking Pex5p-S507/523D (Pex5p(2D)) show decreased import of GFP with a PTS1. We show that the binding affinity between a PTS1-protein and Pex5p(2D) is reduced. An in vivo analysis of the effect of the phospho-mimicking mutant on PTS1-proteins revealed that import of most, but not all, cargos is affected. The physiological effect of the phosphomimetic mutations correlates with the binding affinity of the corresponding extended PTS1-sequences. Thus, we report a novel Pex5p phosphorylation-dependent mechanism for regulating PTS1-protein import into peroxisomes. In a broader view, this suggests that posttranslational modifications can function in fine-tuning the peroxisomal protein composition and, thus, cellular metabolism.
@article{fischerPhosphorylationReceptorProtein2023a,
abstract = {Peroxisomes are organelles with vital functions in metabolism and their dysfunction is associated with human diseases. To fulfill their multiple roles, peroxisomes import nuclear-encoded matrix proteins, most carrying a peroxisomal targeting signal (PTS) 1. The receptor Pex5p recruits PTS1-proteins for import into peroxisomes; whether and how this process is posttranslationally regulated is unknown. Here, we identify 22 phosphorylation sites of Pex5p. Yeast cells expressing phospho-mimicking Pex5p-S507/523D (Pex5p(2D)) show decreased import of GFP with a PTS1. We show that the binding affinity between a PTS1-protein and Pex5p(2D) is reduced. An in vivo analysis of the effect of the phospho-mimicking mutant on PTS1-proteins revealed that import of most, but not all, cargos is affected. The physiological effect of the phosphomimetic mutations correlates with the binding affinity of the corresponding extended PTS1-sequences. Thus, we report a novel Pex5p phosphorylation-dependent mechanism for regulating PTS1-protein import into peroxisomes. In a broader view, this suggests that posttranslational modifications can function in fine-tuning the peroxisomal protein composition and, thus, cellular metabolism.},
added-at = {2024-05-17T13:01:35.000+0200},
address = {Germany},
author = {Fischer, Sven and B{\"u}rgi, J{\'e}r{\^o}me and {Gabay-Maskit}, Shiran and Maier, Renate and Mastalski, Thomas and Yifrach, Eden and {Obarska-Kosinska}, Agnieszka and Rudowitz, Markus and Erdmann, Ralf and Platta, Harald W. and Wilmanns, Matthias and Schuldiner, Maya and Zalckvar, Einat and Oeljeklaus, Silke and Drepper, Friedel and Warscheid, Bettina},
biburl = {https://www.bibsonomy.org/bibtex/22ea2c68b6aa4fe8caac249d7bed07ff2/warscheidlab},
copyright = {{\copyright} 2022 the author(s), published by De Gruyter, Berlin/Boston.},
doi = {10.1515/hsz-2022-0168},
interhash = {fa8b255a7b34369a6c4ba43e9a3c828e},
intrahash = {2ea2c68b6aa4fe8caac249d7bed07ff2},
issn = {1437-4315 1431-6730},
journal = {Biological chemistry},
keywords = {*Peroxisomes/metabolism,*Receptors 1 Cytoplasmic Nuclear/metabolism,Carrier Proteins/metabolism,high-content Receptor/metabolism,Pex5p Signal TPR Transport,Saccharomyces and cerevisiae/metabolism,to_read domain,Phosphorylation,posttranslational localization,Protein modification,protein screen,Humans,mass spectrometry,Peroxisome-Targeting},
langid = {english},
month = feb,
number = {2-3},
pages = {135--155},
pmcid = {PMC9929924},
pmid = {36122347},
timestamp = {2024-05-17T13:01:35.000+0200},
title = {Phosphorylation of the Receptor Protein {{Pex5p}} Modulates Import of Proteins into Peroxisomes.},
volume = 404,
year = 2023
}