Measuring ER stress and the unfolded protein response using mammalian tissue culture system
Reads0
Chats0
TLDR
The current methods to measure ER stress levels, UPR activation, and subsequent pathways in mammalian cells will assist in understanding the UPR and its contribution to ER stress-related disorders such as diabetes and neurodegeneration.Abstract:
The endoplasmic reticulum (ER) functions to properly fold and process secreted and transmembrane proteins. Environmental and genetic factors that disrupt ER function cause an accumulation of misfolded and unfolded proteins in the ER lumen, a condition termed ER stress. ER stress activates a signaling network called the Unfolded Protein Response (UPR) to alleviate this stress and restore ER homeostasis, promoting cell survival and adaptation. However, under unresolvable ER stress conditions, the UPR promotes apoptosis. Here, we discuss the current methods to measure ER stress levels, UPR activation, and subsequent pathways in mammalian cells. These methods will assist us in understanding the UPR and its contribution to ER stress-related disorders such as diabetes and neurodegeneration.read more
Citations
More filters
Journal ArticleDOI
THE JOURNAL OF BIOLOGICAL CHEMISTRY: The Biochemical Behavior of LeadL I. Influence of Calcium, Phosphorus, and Vitamin D on Lead in Blood and Bone
Journal ArticleDOI
A Multiplexed Single-Cell CRISPR Screening Platform Enables Systematic Dissection of the Unfolded Protein Response.
Britt Adamson,Thomas M. Norman,Marco Jost,Min Y. Cho,James K. Nuñez,Yuwen Chen,Jacqueline E. Villalta,Luke A. Gilbert,Max A. Horlbeck,Marco Y. Hein,Ryan A. Pak,Andrew N. Gray,Carol A. Gross,Atray Dixit,Oren Parnas,Aviv Regev,Jonathan S. Weissman +16 more
TL;DR: Perturb-seq as mentioned in this paper combines droplet-based single-cell RNA-seq with a strategy for barcoding CRISPR-mediated perturbations, allowing many perturbation to be profiled in pooled format.
A Multiplexed Single-Cell CRISPR Screening Platform Enables Systematic Dissection of the Unfolded Protein Response
Britt Adamson,Thomas M. Norman,Marco Jost,Min Y. Cho,James K. Nuñez,Yuwen Chen,Jacqueline E. Villalta,Luke A. Gilbert,Max A. Horlbeck,Marco Y. Hein,Ryan A. Pak,Andrew N. Gray,Carol A. Gross,Oren Parnas,Jonathan S. Weissman,Atray Dixit,Aviv Regev +16 more
TL;DR: Insight is provided into how the three sensors of ER homeostasis monitor distinct types of stress and the ability of Perturb-seq to dissect complex cellular responses are highlighted.
Journal ArticleDOI
Endoplasmic Reticulum Stress signalling - from basic mechanisms to clinical applications
Aitor Almanza,Antonio Carlesso,Chetan Chintha,Stuart Creedican,Dimitrios Doultsinos,Brian Leuzzi,Andreia Luís,Nicole McCarthy,Luigi Montibeller,Sanket More,Alexandra Papaioannou,Franziska Püschel,Maria Livia Sassano,Josip Skoko,Patrizia Agostinis,Jackie de Belleroche,Leif A. Eriksson,Simone Fulda,Adrienne M. Gorman,Sandra Healy,Andrey V. Kozlov,Cristina Muñoz-Pinedo,Markus Rehm,Eric Chevet,Afshin Samali +24 more
TL;DR: This review provides a synthesis of intracellular ER signalling revolving around proteostasis and the UPR, its impact on other organelles and cellular behaviour, its multifaceted and dynamic response to stress and its role in physiology.
Journal ArticleDOI
IRE1: ER stress sensor and cell fate executor
Yani Chen,Federica Brandizzi +1 more
TL;DR: An updated scenario of the IRE1 signaling model is provided, a discussion of emerging IRE 1 sensing mechanisms is discussed, features among species are compared, and exciting future directions in UPR research are outlined.
References
More filters
Journal ArticleDOI
Signal integration in the endoplasmic reticulum unfolded protein response
David Ron,Peter Walter +1 more
TL;DR: Together, at least three mechanistically distinct arms of the UPR regulate the expression of numerous genes that function within the secretory pathway but also affect broad aspects of cell fate and the metabolism of proteins, amino acids and lipids.
Journal ArticleDOI
Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B.
TL;DR: It is shown that agents which prevent the activation of both MAPKAP kinase-1 and p70S6k by insulin in vivo do not block the phosphorylation and inhibition of GSK3, and it is demonstrated that PKB is the product of the proto-oncogene protein kinase B (PKB, also known as Akt/RAC).
Journal ArticleDOI
XBP1 mRNA Is Induced by ATF6 and Spliced by IRE1 in Response to ER Stress to Produce a Highly Active Transcription Factor
TL;DR: The transcription factor XBP1, a target of ATF6, is identified as a mammalian substrate of such an unconventional mRNA splicing system and it is shown that only the spliced form of X BP1 can activate the UPR efficiently.
Journal ArticleDOI
Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase
TL;DR: The cloning of perk is described, a gene encoding a type I transmembrane ER-resident protein that contains a protein-kinase domain most similar to that of the known eIF2α kinases, PKR and HRI that implicate PERK in a signalling pathway that attenuates protein translation in response to ER stress.
Journal ArticleDOI
Regulated Translation Initiation Controls Stress-Induced Gene Expression in Mammalian Cells
Heather P. Harding,Isabel Novoa,Yuhong Zhang,Huiqing Zeng,Ronald C. Wek,Matthieu Schapira,David Ron +6 more
TL;DR: Protein kinases that phosphorylate the alpha subunit of eukaryotic initiation factor 2 (eIF2alpha) are activated in stressed cells and negatively regulate protein synthesis, resulting in the induction of the downstream gene CHOP (GADD153).
Related Papers (5)
The Unfolded Protein Response: From Stress Pathway to Homeostatic Regulation
Peter Walter,David Ron +1 more
Signal integration in the endoplasmic reticulum unfolded protein response
David Ron,Peter Walter +1 more