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Impairment of RPN4, a transcription factor, induces ER stress and lipid abnormality in Saccharomyces cerevisiae

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Abstract

Accumulation of misfolded/unfolded proteins in the endoplasmic reticulum (ER) induces ER stress. The transcription factor RPN4 {“Regulatory Particle Non-ATPase”} regulates protein homeostasis by degrading proteins that elude proper folding or assembly via the proteasomal degradation pathway. Here, we studied the lipid alterations exerted by Saccharomyces cerevisiae to mitigate (ER) stress during adaptive responses in rpn4∆ cells. The loss of RPN4-induced ER stress increased phospholipid synthesis, leading to altered membrane structures and accumulation of neutral lipids, causing an increase in lipid droplets (LDs). There was a significant upregulation of genes involved in neutral lipid and membrane lipid synthesis in rpn4∆ cells. Overexpression of RPN4 restored the defects caused by rpn4∆ cells. Thus, our study provides new insight that RPN4 impacts lipid homeostasis.

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All data generated or analyzed during this study are included in this published article (and its supplementary information files).

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Acknowledgements

The author NB was supported by a fellowship (IF150823) from DST-INSPIRE, New Delhi, India. We also thank the UGC-BSR grant for providing financial assistance. We thank DST-FIST for providing infrastructure in the Department of Biochemistry and DST-PURSE for providing the confocal facility at Bharathidasan University. We sincerely thank Prof. J.W. Hanna (Department of Pathology, Brigham and Women’s Hospital and Harvard Medical school, Boston) USA, for gifting us the mutant strain. We thank Prof. Shigeo Murata (Laboratory of Protein Metabolism, The University of Tokyo, Japan) and Prof. Anne Skaja Robinson (Department of Chemical and Biomolecular Engineering, Tulane University, USA) for gifting the overexpression plasmid pYES2-RPN4 and UPRE-GFP. We thank Prof. Jeffrey L. Brodsky (Department of Cell Biology, University of Pittsburgh, USA) for providing Kar2 antibodies. We thank Prof. Ram Rajasekharan, Head Department of Microbiology, Central University of Tamil Nadu, Thiruvarur, India, for providing reagents for this study.

Funding

This work is supported by the UGC-BSR faculty fellowship grant [F4-5(11)/2019(BSR)],New Delhi,India and INSPIRE fellowship [IF150823],DST,New Delhi, India.

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Authors and Affiliations

  1. Biomembrane Lab, Department of Biochemistry, School of Life Sciences, Bharathidasan University, Trichy, Tamil Nadu, 620 024, India

    Bhanupriya Nagaraj, Antonisamy William James, Arul Mathivanan & Vasanthi Nachiappan

  2. Departments of Medicine and Cancer Biology, College of Medicine & Life Sciences, Toledo, USA

    Antonisamy William James

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  1. Bhanupriya Nagaraj
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  2. Antonisamy William James
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  4. Vasanthi Nachiappan
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Conceived and designed the experiments: BN, AWJ, and VN. Experiments performed: BN. Analyzing data: BN and VN. Drafted the manuscript: BN, AM, and VN; all the authors reviewed the manuscript.

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Correspondence to Vasanthi Nachiappan.

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Nagaraj, B., James, A.W., Mathivanan, A. et al. Impairment of RPN4, a transcription factor, induces ER stress and lipid abnormality in Saccharomyces cerevisiae. Mol Cell Biochem 478, 2127–2139 (2023). https://doi.org/10.1007/s11010-022-04623-w

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