de Lange Eline M F, Mol Frank N, van der Klei Ida J, Vlijm Rifka
Molecular Cell Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 7, 9747 AG Groningen, the Netherlands.
Molecular Biophysics, Zernike Institute for Advanced Materials, University of Groningen, Nijenborgh 4, 9747 AG Groningen, the Netherlands.
iScience. 2024 Jul 8;27(8):110481. doi: 10.1016/j.isci.2024.110481. eCollection 2024 Aug 16.
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 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.
过氧化物酶体是具有重要代谢功能的动态细胞器。酵母Pex3是一种多功能膜蛋白，通过与特定过程因子相互作用，辅助过氧化物酶体的生物发生、遗传和降解（过氧化物酶体自噬）。利用多色（活细胞）受激发射损耗（STED）纳米显微镜，我们研究了Pex3及其结合伴侣在……中的定位。与共聚焦显微镜不同，STED能够分辨微小过氧化物酶体的膜，从而能够准确测量所有Pex3标记的过氧化物酶体的大小。我们在过氧化物酶体抑制和诱导条件下以及过氧化物酶体自噬过程中定位了Pex3及其结合伴侣。对Pex3和过氧化物酶体自噬受体Atg30的深入定量分析显示了它们（共）定位的动态变化。Atg30的一个显著反应是其位置的转变，从在增殖条件下夹在聚集的过氧化物酶体之间，到在过氧化物酶体自噬诱导时位于过氧化物酶体簇的胞质暴露部分。总之，我们表明STED能够表征酵母细胞中过氧化物酶体蛋白定位的动态变化。
