Department of Genetics, Graduate School of Medicine, Osaka University, Osaka, Japan.
Department of Intracellular Membrane Dynamics, Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan.
J Cell Biol. 2023 Dec 4;222(12). doi: 10.1083/jcb.202204048. Epub 2023 Oct 6.
Membrane rupture of lysosomes results in leakage of their contents, which is harmful to cells. Recent studies have reported that several systems contribute to the repair or elimination of damaged lysosomes. Lysophagy is a type of selective autophagy that plays a crucial role in the lysosomal damage response. Because multiple pathways are involved in this response, an assay that specifically evaluates lysophagy is needed. Here, we developed the TMEM192-mKeima probe to evaluate lysophagy. By comparing the use of this probe with the conventional galectin-3 assay, we showed that this probe is more specific to lysophagy. Using TMEM192-mKeima, we showed that TFEB and p62 are important for the lysosomal damage response but not for lysophagy, although they have previously been considered to be involved in lysophagy. We further investigated the initial steps in lysophagy and identified UBE2L3, UBE2N, TRIM10, 16, and 27 as factors involved in it. Our results demonstrate that the TMEM192-mKeima probe is a useful tool for investigating lysophagy.
溶酶体膜破裂导致其内容物泄漏，这对细胞是有害的。最近的研究报告称，有几个系统有助于受损溶酶体的修复或清除。溶酶体自噬是一种选择性自噬，在溶酶体损伤反应中起着至关重要的作用。由于该反应涉及多个途径，因此需要一种专门评估溶酶体自噬的测定方法。在这里，我们开发了 TMEM192-mKeima 探针来评估溶酶体自噬。通过比较使用该探针与传统半乳糖凝集素-3 测定法，我们表明该探针对溶酶体自噬的特异性更高。使用 TMEM192-mKeima，我们表明 TFEB 和 p62 对溶酶体损伤反应很重要，但对溶酶体自噬不重要，尽管它们以前被认为参与了溶酶体自噬。我们进一步研究了溶酶体自噬的初始步骤，确定了 UBE2L3、UBE2N、TRIM10、16 和 27 是参与其中的因素。我们的结果表明，TMEM192-mKeima 探针是研究溶酶体自噬的有用工具。
