Sqstm1–GFP knock-in mice reveal dynamic actions of Sqstm1 during autophagy and under stress conditions in living cells
Sqstm1 serves as a signaling hub and receptor for selective autophagy. Consequently, dysregulation of Sqstm1 causes imbalances in signaling pathways and disrupts proteostasis, thereby contributing to the development of human diseases. Environmental stresses influence the level of Sqstm1 by altering...
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The Company of Biologists
2015
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4712822/ |
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pubmed-4712822 |
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pubmed-47128222016-02-05 Sqstm1–GFP knock-in mice reveal dynamic actions of Sqstm1 during autophagy and under stress conditions in living cells Eino, Atsushi Kageyama, Shun Uemura, Takefumi Annoh, Hiromichi Saito, Tetsuya Narita, Ichiei Waguri, Satoshi Komatsu, Masaaki Tools and Techniques Sqstm1 serves as a signaling hub and receptor for selective autophagy. Consequently, dysregulation of Sqstm1 causes imbalances in signaling pathways and disrupts proteostasis, thereby contributing to the development of human diseases. Environmental stresses influence the level of Sqstm1 by altering its expression and/or autophagic degradation, and also changes the localization of Sqstm1, making it difficult to elucidate the actions and roles of this protein. In this study, we developed knock-in mice expressing Sqstm1 fused to GFP (Sqstm1-GFPKI/+). Using these Sqstm1-GFPKI/+ mice, we revealed for the first time the dynamics of endogenous Sqstm1 in living cells. Sqstm1–GFP was translocated to a restricted area of LC3-positive structures, which primarily correspond to the inside of autophagosomes, and then degraded. Moreover, exposure to arsenite induced expression of Sqstm1–GFP, followed by accumulation of the fusion protein in large aggregates that were degraded by autophagy. Furthermore, suppression of autophagy in Sqstm1-GFPKI/+ mouse livers caused accumulation of Sqstm1–GFP and formation of GFP-positive aggregate structures, leading to severe hepatic failure. These results indicate that Sqstm1-GFPKI/+ mice are a useful tool for analyzing Sqstm1 in living cells and intact animals. The Company of Biologists 2015-12-01 /pmc/articles/PMC4712822/ /pubmed/26483381 http://dx.doi.org/10.1242/jcs.180174 Text en © 2015. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Eino, Atsushi Kageyama, Shun Uemura, Takefumi Annoh, Hiromichi Saito, Tetsuya Narita, Ichiei Waguri, Satoshi Komatsu, Masaaki |
| spellingShingle |
Eino, Atsushi Kageyama, Shun Uemura, Takefumi Annoh, Hiromichi Saito, Tetsuya Narita, Ichiei Waguri, Satoshi Komatsu, Masaaki Sqstm1–GFP knock-in mice reveal dynamic actions of Sqstm1 during autophagy and under stress conditions in living cells |
| author_facet |
Eino, Atsushi Kageyama, Shun Uemura, Takefumi Annoh, Hiromichi Saito, Tetsuya Narita, Ichiei Waguri, Satoshi Komatsu, Masaaki |
| author_sort |
Eino, Atsushi |
| title |
Sqstm1–GFP knock-in mice reveal dynamic actions of Sqstm1 during autophagy and under stress conditions in living cells |
| title_short |
Sqstm1–GFP knock-in mice reveal dynamic actions of Sqstm1 during autophagy and under stress conditions in living cells |
| title_full |
Sqstm1–GFP knock-in mice reveal dynamic actions of Sqstm1 during autophagy and under stress conditions in living cells |
| title_fullStr |
Sqstm1–GFP knock-in mice reveal dynamic actions of Sqstm1 during autophagy and under stress conditions in living cells |
| title_full_unstemmed |
Sqstm1–GFP knock-in mice reveal dynamic actions of Sqstm1 during autophagy and under stress conditions in living cells |
| title_sort |
sqstm1–gfp knock-in mice reveal dynamic actions of sqstm1 during autophagy and under stress conditions in living cells |
| description |
Sqstm1 serves as a signaling hub and receptor for selective autophagy. Consequently, dysregulation of Sqstm1 causes imbalances in signaling pathways and disrupts proteostasis, thereby contributing to the development of human diseases. Environmental stresses influence the level of Sqstm1 by altering its expression and/or autophagic degradation, and also changes the localization of Sqstm1, making it difficult to elucidate the actions and roles of this protein. In this study, we developed knock-in mice expressing Sqstm1 fused to GFP (Sqstm1-GFPKI/+). Using these Sqstm1-GFPKI/+ mice, we revealed for the first time the dynamics of endogenous Sqstm1 in living cells. Sqstm1–GFP was translocated to a restricted area of LC3-positive structures, which primarily correspond to the inside of autophagosomes, and then degraded. Moreover, exposure to arsenite induced expression of Sqstm1–GFP, followed by accumulation of the fusion protein in large aggregates that were degraded by autophagy. Furthermore, suppression of autophagy in Sqstm1-GFPKI/+ mouse livers caused accumulation of Sqstm1–GFP and formation of GFP-positive aggregate structures, leading to severe hepatic failure. These results indicate that Sqstm1-GFPKI/+ mice are a useful tool for analyzing Sqstm1 in living cells and intact animals. |
| publisher |
The Company of Biologists |
| publishDate |
2015 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4712822/ |
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1613524028771794944 |