Targeted disruption of GAK stagnates autophagic flux by disturbing lysosomal dynamics
| Autor: | Hirotsugu Hino, Mayumi Tokuhisa, Keisuke Miyazawa, Masaki Hiramoto, Naoharu Takano, Jun Takemura, Hiroko Kokuba, Masaya Miyazaki, Hiromi Kazama |
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| Rok vydání: | 2021 |
| Předmět: |
autophagy
autophagic lysosome reformation Protein Serine-Threonine Kinases Endocytosis Rho-associated protein kinase Lysosome Genetics medicine Humans ROCK1 Protein kinase A Cyclin rho-Associated Kinases autophagosome-lysosome fusion actomyosin Chemistry cyclin G-associated kinase Autophagy Autophagosomes Intracellular Signaling Peptides and Proteins Articles General Medicine Cell cycle Cell biology medicine.anatomical_structure A549 Cells Lysosomes Flux (metabolism) |
| Zdroj: | International Journal of Molecular Medicine |
| ISSN: | 1791-244X 1107-3756 |
| DOI: | 10.3892/ijmm.2021.5028 |
| Popis: | The autophagy‑lysosome system allows cells to adapt to environmental changes by regulating the degradation and recycling of cellular components, and to maintain homeostasis by removing aggregated proteins and defective organelles. Cyclin G‑associated kinase (GAK) is involved in the regulation of clathrin‑dependent endocytosis and cell cycle progression. In addition, a single nucleotide polymorphism at the GAK locus has been reported as a risk factor for Parkinson's disease. However, the roles of GAK in the autophagy‑lysosome system are not completely understood, thus the present study aimed to clarify this. In the present study, under genetic disruption or chemical inhibition of GAK, analyzing autophagic flux and observing morphological changes of autophagosomes and autolysosomes revealed that GAK controlled lysosomal dynamics via actomyosin regulation, resulting in a steady progression of autophagy. GAK knockout (KO) in A549 cells impaired autophagosome‑lysosome fusion and autophagic lysosome reformation, which resulted in the accumulation of enlarged autophagosomes and autolysosomes during prolonged starvation. The stagnation of autophagic flux accompanied by these phenomena was also observed with the addition of a GAK inhibitor. Furthermore, the addition of Rho‑associated protein kinase (ROCK) inhibitor or ROCK1 knockdown mitigated GAK KO‑mediated effects. The results suggested a vital role of GAK in controlling lysosomal dynamics via maintaining lysosomal homeostasis during autophagy. |
| Databáze: | OpenAIRE |
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