Abstract
RNF5 is implicated in ERAD and in negative regulation of macroautophagy/autophagy. To better understand the function of RNF-5 under ER-stress conditions, we studied the ability of Caenorhabditis elegans rnf-5(tm794) mutant animals to cope with stress in the background of impaired UPR machinery. We demonstrate that downregulation of RNF-5 decreased sensitivity to tunicamycin both in wild type and in an ire-1 mutant. Double-mutant rnf-5;ire-1 animals showed increased starvation resistance and extended lifespan when compared to the ire-1 mutant. This partial rescue of ire-1 required functional autophagy. Downregulation of RNF-5 rescued ER maturation defects and protein secretion of a DAF-28::GFP intestinal reporter in the ire-1 background. Proteomics and functional studies revealed an increase in lysosomal protease levels, in the frequency of intestinal lysosomes, and in lysosomal protease activity in rnf-5(tm794) animals. Together, these data suggest that RNF-5 is a negative regulator of ER stress, and that inactivation of RNF-5 promotes IRE-1-independent elevation of ER capacity.
| Original language | English |
|---|---|
| Pages (from-to) | 2401-2414 |
| Number of pages | 14 |
| Journal | Autophagy |
| Volume | 17 |
| Issue number | 9 |
| Early online date | 15 Oct 2020 |
| DOIs | |
| State | Published - Sep 2021 |
Bibliographical note
Funding Information:We thank WormBase for C. elegans genetic annotation. Several strains were provided by the Caenorhabditis Genetics Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440) and Dr. Shohei Mitani, National Bioresource Project for the nematode, Tokyo Women’s Medical University School of Medicine, Japan. We thank Prof. Dieter A. Wolf from the Sanford Burnham Prebys Medical Discovery Institute for advices on proteomics analysis and comments on the manuscript. We thank Prof. Peter Naredi (Umea University, Sweden) for the DAF-28::GFP expressing strain. We thank Sharon Sheffy-Levin for sample preparation for the proteomics analysis and initial hit analysis and Hanna Grobe for writing and optimizing the ImageJ Macro for lysosome analysis.
Funding Information:
This research was supported by the Israel Science Foundation (ISF 1878/15 and 2122/19) and the Israel Cancer Research Fund PG-14-101 (to L.B.), and by the Ministry of Science, Technology & Space, Israel (3-12066) (to S.H-K). We thank WormBase for C. elegans genetic annotation. Several strains were provided by the Caenorhabditis Genetics Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440) and Dr. Shohei Mitani, National Bioresource Project for the nematode, Tokyo Women’s Medical University School of Medicine, Japan. We thank Prof. Dieter A. Wolf from the Sanford Burnham Prebys Medical Discovery Institute for advices on proteomics analysis and comments on the manuscript. We thank Prof. Peter Naredi (Umea University, Sweden) for the DAF-28::GFP expressing strain. We thank Sharon Sheffy-Levin for sample preparation for the proteomics analysis and initial hit analysis and Hanna Grobe for writing and optimizing the ImageJ Macro for lysosome analysis.
Publisher Copyright:
© 2020 Informa UK Limited, trading as Taylor & Francis Group.
Keywords
- IRE1
- RMA1
- RNF5
- autophagy, C. elegans
- endoplasmic reticulum
- lysosome