Related Experiment Video
Updated: Nov 8, 2025

Assays for the Degradation of Misfolded Proteins in Cells
Published on: August 28, 2016
Tolerance to nascent protein misfolding stress requires fine-tuning of the cAMP/PKA pathway
Paraskevi Kritsiligkou1, Karol Nowicki-Osuch1, Zorana Carter1
1Faculty of Biology, Medicine and Health, The University of Manchester, Manchester, UK.
Abstract:
Protein aggregation is the abnormal association of misfolded proteins into larger, often insoluble structures that can be toxic during aging and in protein aggregation-associated diseases. Previous research has established a role for the cytosolic Tsa1 peroxiredoxin in responding to protein misfolding stress. Tsa1 is also known to downregulate the cAMP/protein kinase A (PKA) pathway as part of the response to hydrogen peroxide stress. However, whether the cAMP/PKA pathway is involved in protein misfolding stress is not known. Using transcriptomics, we examined the response to protein misfolding stress and found upregulation of numerous stress gene functions and downregulation of many genes related to protein synthesis and other growth-related processes consistent with the well-characterized environmental stress response. The scope of the transcriptional response is largely similar in wild-type and tsa1 mutant strains, but the magnitude is dampened in the strain lacking Tsa1. We identified a direct protein interaction between Tsa1 and the Bcy1 regulatory subunit of PKA that is present under normal growth conditions and explains the observed differences in gene expression profiles. This interaction is increased in a redox-dependent manner in response to nascent protein misfolding, via Tsa1-mediated oxidation of Bcy1. Oxidation of Bcy1 causes a reduction in cAMP binding by Bcy1, which dampens PKA pathway activity, leading to a targeted reprogramming of gene expression. Redox regulation of the regulatory subunit of PKA provides a mechanism to mitigate the toxic consequences of protein misfolding stress that is distinct to stress caused by exogenous sources of reactive oxygen species.
Insights
The cytosolic Tsa1 peroxiredoxin interacts with the Bcy1 subunit of protein kinase A (PKA) to regulate gene expression during protein misfolding stress. This redox-dependent interaction dampens PKA activity, mitigating cellular damage.
Area of Science:
- Cellular Biology
- Biochemistry
- Molecular Biology
Background:
- Protein aggregation is implicated in aging and disease.
- Cytosolic Tsa1 peroxiredoxin responds to protein misfolding stress.
- Tsa1 downregulates the cAMP/protein kinase A (PKA) pathway during hydrogen peroxide stress.
Purpose of the Study:
- To investigate the involvement of the cAMP/PKA pathway in protein misfolding stress.
- To elucidate the role of Tsa1 in this stress response.
- To identify molecular mechanisms linking Tsa1 and PKA.
Main Methods:
- Transcriptomics to analyze gene expression changes.
- Protein interaction studies to identify direct binding partners.
- Redox-dependent assays to assess molecular modifications.
Main Results:
- Protein misfolding stress induces a transcriptional response involving stress genes and growth-related processes.
- Tsa1 modulates the magnitude of this transcriptional response.
- Tsa1 directly interacts with Bcy1, the regulatory subunit of PKA.
- This interaction is redox-dependent, increasing upon protein misfolding.
- Oxidation of Bcy1 by Tsa1 reduces cAMP binding, dampening PKA activity.
Conclusions:
- Redox regulation of Bcy1 by Tsa1 is a novel mechanism to control PKA activity during protein misfolding stress.
- This pathway reprogramming mitigates the toxic effects of protein aggregation.
- This stress response is distinct from responses to exogenous reactive oxygen species.
Related Concept Videos
Bacterial Protein Maturation
The Unfolded Protein Response
Molecular Chaperones and Protein Folding
The...
Molecular Chaperones and Protein Folding
Protein Folding
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
Protein Folding

