TY - JOUR
T1 - Inactivation of the Caenorhabditis elegans RNF-5 E3 ligase promotes IRE-1-independent ER functions
AU - Adir, Orit
AU - Bening-Abu-Shach, Ulrike
AU - Arbib, Shir
AU - Henis-Korenblit, Sivan
AU - Broday, Limor
N1 - 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.
Publisher Copyright:
© 2020 Informa UK Limited, trading as Taylor & Francis Group.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/10/15
Y1 - 2020/10/15
N2 - 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.
AB - 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.
KW - autophagy, C. elegans
KW - endoplasmic reticulum
KW - IRE1
KW - lysosome
KW - RMA1
KW - RNF5
UR - http://www.scopus.com/inward/record.url?scp=85092558441&partnerID=8YFLogxK
U2 - 10.1080/15548627.2020.1827778
DO - 10.1080/15548627.2020.1827778
M3 - מאמר
C2 - 32981418
AN - SCOPUS:85092558441
VL - 17
SP - 2401
EP - 2414
JO - Autophagy
JF - Autophagy
SN - 1554-8627
IS - 9
ER -