Inhibitory effect of mTOR activator MHY1485 on autophagy: suppression of lysosomal fusion
Open accessAbstract
Autophagy is a major degradative process responsible for the disposal of cytoplasmic proteins and dysfunctional organelles via the lysosomal pathway. During the autophagic process, cells form double-membraned vesicles called autophagosomes that sequester disposable materials in the cytoplasm and finally fuse with lysosomes. In the present study, we investigated the inhibition of autophagy by a synthesized compound, MHY1485, in a culture system by using Ac2F rat hepatocytes. Autophagic flux was measured to evaluate the autophagic activity. Autophagosomes were visualized in Ac2F cells transfected with AdGFP-LC3 by live-cell confocal microscopy. In addition, activity of mTOR, a major regulatory protein of autophagy, was assessed by western blot and docking simulation using AutoDock 4.2. In the result, treatment with MHY1485 suppressed the basal autophagic flux, and this inhibitory effect was clearly confirmed in cells under starvation, a strong physiological inducer of autophagy. The levels of p62 and beclin-1 did not show significant change after treatment with MHY1485. Decreased co-localization of autophagosomes and lysosomes in confocal microscopic images revealed the inhibitory effect of MHY1485 on lysosomal fusion during starvation-induced autophagy. These effects of MHY1485 led to the accumulation of LC3II and enlargement of the autophagosomes in a dose- and time-dependent manner. Furthermore, MHY1485 induced mTOR activation and correspondingly showed a higher docking score than PP242, a well-known ATP-competitive mTOR inhibitor, in docking simulation. In conclusion, MHY1485 has an inhibitory effect on the autophagic process by inhibition of fusion between autophagosomes and lysosomes leading to the accumulation of LC3II protein and enlarged autophagosomes. MHY1485 also induces mTOR activity, providing a possibility for another regulatory mechanism of autophagy by the MHY compound. The significance of this study is the finding of a novel inhibitor of autophagy with an mTOR activating effect.
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mTORC1 signaling may disrupt autophagy, which in turn may affect mitochondrial remodeling.
“There are also data showing mTORC1 signaling may disrupt autophagy, which in turn may affect mitochondrial remodeling (Choi et al., 2012).”
From Skeletal Muscle Ribosome and Mitochondrial Biogenesis in Response to Different Exercise Training Modalities (2021) · cites this paper as Choi et al., 2012
The Discussion (page unavailable) reports that MHY1485 inhibited autophagosome–lysosome fusion and activated mTOR in rat hepatocytes, but says further study is needed to establish whether the mTOR activation regulates autophagy. The paper does not address mitochondrial remodeling, so it supports the autophagy-disruption aspect but not the proposed downstream effect. (LLM, conf=0.88)
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- Authors
- Choi, Y. J., Park, Y. J., Park, J. Y., Jeong, H. O., Kim, D. H., Ha, Y. M., Kim, J. M., Song, Y. M., Heo, H.-S., Yu, B. P., Chun, P., Moon, H. R., & Chung, H. Y.
- Affiliations
- Pusan National University (KR)
- Korea Research Institute of Bioscience and Biotechnology (KR)
- The University of Texas at San Antonio Health Science Center (US)
- Inje University (KR)
- Corresponding author
- Moon, Hyung Ryong
- Chung, Hae Young
- ORCID
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2012-08-22
- Volume
- 7
- Issue
- 8
- Pages
- e43418
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC3425474
- OpenAlex
- W1985820389
- Cited by
- 192
- References
- 47
- Citations by year
- 2026: 18 · 2025: 13 · 2024: 28 · 2023: 16 · 2022: 14 · 2021: 16 · 2020: 17 · 2019: 10 · 2018: 17 · 2017: 15 · 2016: 10 · 2015: 9 · 2014: 6 · 2013: 3
- FWCI
- 4.29
- Type
- journal article
- Language
- en
- Update notices
- correction (2013-01-29)
- Open access
- gold
- doi.org
- Locations
- PLoS ONE (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- KRIBB Repository (submittedVersion)
- Funders
- National Research Foundation
- National Research Foundation of Korea (20090083538, No. 2009-0083538)
- Topics
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Calcium signaling and nucleotide metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › PI3K/AKT/mTOR signaling in cancer
- Keywords
- MHY1485, autophagy inhibition, mTOR activation, lysosome fusion, autophagosome-lysosome fusion, starvation-induced autophagy, autophagic flux, hepatocytes, p62, Beclin-1, mTOR inhibitors
- MeSH terms
- Animals, Autophagy, Liver, Lysosomes, Microtubule-Associated Proteins, Morpholines, Protein Conformation, Triazines, Vacuoles, Rats, TOR Serine-Threonine Kinases, Molecular Docking Simulation
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