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Altered Lysosomal Function Manipulates Cellular Biosynthetic Capacity By Remodeling Intracellular Cholesterol Distribution

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Abstract

Lysosomes are known to influence cholesterol trafficking into endoplasmic reticulum (ER) membranes. Though intracellular cholesterol levels are known to influence the lipid biosynthetic responses in ER, the specific effects of lysosomal modulation on these outcomes is not known. To demonstrate this, C2C12 cells were treated with chloroquine, a lysosomotropic agent, and its effects on cellular biosynthetic capacity, structural and functional status of ER was determined. In addition to its known effects on autophagy reduction, chloroquine treatment induced accumulation of total cellular lipid and ER-specific cholesterol content. It was also observed that chloroquine caused an increase in smooth-ER content with defects in overall protein turnover. Further, since ER and mitochondria function in close association through ER membrane contact sites, it is likely that lysosomal modulation also brings about associated changes in mitochondria. In this regard, we found that chloroquine reduces mitochondrial membrane potential and mitochondrial dynamics. Collectively, the differential biosynthetic response of rise in lipid content, but not protein content, cannot be accounted by merely considering that chloroquine induced suppression of autophagy causes defects in organelle function. In this defective autophagy scenario, both biosynthetic responses such as lipid and protein synthesis are expected to be reduced rather than only the latter, as observed with chloroquine. These findings suggest that cholesterol trafficking/distribution within cellular organelles could act as an intracellular mediator of differential biosynthetic remodelling in interconnected organelles.

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Acknowledgements

This work was funded by Science and Engineering Research Board (CRG/2020/003399), India. We thank Indian Council of Medical Research (File No. 2017-3862/CMB-BMS) and Council of Scientific and Industrial Research (File No. 09/468(0489)/2015-EMR-I) for awarding Senior Research Fellowship to S.R. We thank Department of Science and Technology, India- Promotion of University Research and Scientific Excellence (PURSE), Fund for Improvement of S&T Infrastructure in Universities and Higher Educational Institutions (FIST) and Department of Biotechnology, India- Boost to University Interdisciplinary Life science Departments for Education and Research (BUILDER) (BT/PR12153/INF/22/200/2014) for chemiluminiscence, confocal microscope and HPLC instrumentation respectively.

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  1. Centre for Biotechnology, Anna University, Chennai, 600025, India

    Sudarshana Rajasekaran, Anand Ramaian Santhaseela, Shamila Ragunathan, Sangita Venkataraman & Tamilselvan Jayavelu

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S.R. was involved in Data collection, Data analysis and draft paper preparation, A.R.S. contributed to Data analysis and interpretation, S.R. was involved in Data analysis, S.V. was involved in Data analysis, T.J. was involved in study conception and design, data analysis and paper preparation.

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Correspondence to Tamilselvan Jayavelu.

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Rajasekaran, S., Ramaian Santhaseela, A., Ragunathan, S. et al. Altered Lysosomal Function Manipulates Cellular Biosynthetic Capacity By Remodeling Intracellular Cholesterol Distribution. Cell Biochem Biophys 81, 29–38 (2023). https://doi.org/10.1007/s12013-022-01123-y

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