STED super-resolution microscopy unveils the dynamics of Atg30 on yeast Pex3-labeled peroxisomes

• Published in: iScience 2024-08, Vol.27 (8), p.110481, Article 110481
• Main Authors: de Lange, Eline M.F., Mol, Frank N., van der Klei, Ida J., Vlijm, Rifka
• Format: Article
• Language: English
• Subjects: Biological sciences; Molecular biology; Resolution techniques; Structural biology
• ISSN: 2589-0042; 2589-0042
• DOI: 10.1016/j.isci.2024.110481

Peroxisomes are dynamic organelles with important metabolic functions. Yeast Pex3 is a multifunctional membrane protein aiding in peroxisomal biogenesis, inheritance, and degradation (pexophagy), by interacting with process-specific factors. Using multicolor (live-cell) stimulated emission depletion (STED) nanoscopy, we studied the localization of Pex3 and its binding partners in Hansenula polymorpha. Unlike confocal microscopy, STED allows resolving the membrane of tiny peroxisomes, enabling accurate measurements of the size of all Pex3-labeled peroxisomes. We localized Pex3 and its binding partners at peroxisome-repressing and -inducing conditions and during pexophagy. In-depth quantitative analysis of Pex3 and pexophagy receptor Atg30 showed dynamic changes in their (co)localization. One remarkable response of Atg30 was the shift in position from being sandwiched between clustered peroxisomes at proliferation conditions, to the cytosolically exposed parts of peroxisome clusters upon pexophagy induction. Summarizing, we show that STED allows characterizing dynamics of the localization of peroxisomal proteins in yeast cells. [Display omitted] •STED nanoscopy is feasible in living yeast cells•STED allows characterizing the dynamics of peroxisomal proteins in yeast•Changed colocalization of Pex3 and its binding partners is accurately quantified•Atg30 relocates on the peroxisomal membrane upon pexophagy induction Biological sciences; Molecular biology; Structural biology; Resolution techniques