Identification of mammalian protein quality control factors by high-throughput cellular imaging.
Protein Quality Control (PQC) pathways are essential to maintain the equilibrium between protein folding and the clearance of misfolded proteins. In order to discover novel human PQC factors, we developed a high-content, high-throughput cell-based assay to assess PQC activity. The assay is based on...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3282772?pdf=render |
| _version_ | 1818031562680172544 |
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| author | Gianluca Pegoraro Ty C Voss Scott E Martin Pinar Tuzmen Rajarshi Guha Tom Misteli |
| author_facet | Gianluca Pegoraro Ty C Voss Scott E Martin Pinar Tuzmen Rajarshi Guha Tom Misteli |
| author_sort | Gianluca Pegoraro |
| collection | DOAJ |
| description | Protein Quality Control (PQC) pathways are essential to maintain the equilibrium between protein folding and the clearance of misfolded proteins. In order to discover novel human PQC factors, we developed a high-content, high-throughput cell-based assay to assess PQC activity. The assay is based on a fluorescently tagged, temperature sensitive PQC substrate and measures its degradation relative to a temperature insensitive internal control. In a targeted screen of 1591 siRNA genes involved in the Ubiquitin-Proteasome System (UPS) we identified 25 of the 33 genes encoding for 26S proteasome subunits and discovered several novel PQC factors. An unbiased genome-wide siRNA screen revealed the protein translation machinery, and in particular the EIF3 translation initiation complex, as a novel key modulator of misfolded protein stability. These results represent a comprehensive unbiased survey of human PQC components and establish an experimental tool for the discovery of genes that are required for the degradation of misfolded proteins under conditions of proteotoxic stress. |
| first_indexed | 2024-12-10T05:53:27Z |
| format | Article |
| id | doaj.art-7728615b743e441e85ca77beaa526dcf |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-10T05:53:27Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-7728615b743e441e85ca77beaa526dcf2022-12-22T01:59:59ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0172e3168410.1371/journal.pone.0031684Identification of mammalian protein quality control factors by high-throughput cellular imaging.Gianluca PegoraroTy C VossScott E MartinPinar TuzmenRajarshi GuhaTom MisteliProtein Quality Control (PQC) pathways are essential to maintain the equilibrium between protein folding and the clearance of misfolded proteins. In order to discover novel human PQC factors, we developed a high-content, high-throughput cell-based assay to assess PQC activity. The assay is based on a fluorescently tagged, temperature sensitive PQC substrate and measures its degradation relative to a temperature insensitive internal control. In a targeted screen of 1591 siRNA genes involved in the Ubiquitin-Proteasome System (UPS) we identified 25 of the 33 genes encoding for 26S proteasome subunits and discovered several novel PQC factors. An unbiased genome-wide siRNA screen revealed the protein translation machinery, and in particular the EIF3 translation initiation complex, as a novel key modulator of misfolded protein stability. These results represent a comprehensive unbiased survey of human PQC components and establish an experimental tool for the discovery of genes that are required for the degradation of misfolded proteins under conditions of proteotoxic stress.http://europepmc.org/articles/PMC3282772?pdf=render |
| spellingShingle | Gianluca Pegoraro Ty C Voss Scott E Martin Pinar Tuzmen Rajarshi Guha Tom Misteli Identification of mammalian protein quality control factors by high-throughput cellular imaging. PLoS ONE |
| title | Identification of mammalian protein quality control factors by high-throughput cellular imaging. |
| title_full | Identification of mammalian protein quality control factors by high-throughput cellular imaging. |
| title_fullStr | Identification of mammalian protein quality control factors by high-throughput cellular imaging. |
| title_full_unstemmed | Identification of mammalian protein quality control factors by high-throughput cellular imaging. |
| title_short | Identification of mammalian protein quality control factors by high-throughput cellular imaging. |
| title_sort | identification of mammalian protein quality control factors by high throughput cellular imaging |
| url | http://europepmc.org/articles/PMC3282772?pdf=render |
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