A high-throughput screen of real-time ATP levels in individual cells reveals mechanisms of energy failure.
Insufficient or dysregulated energy metabolism may underlie diverse inherited and degenerative diseases, cancer, and even aging itself. ATP is the central energy carrier in cells, but critical pathways for regulating ATP levels are not systematically understood. We combined a pooled clustered regula...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2018-08-01
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| Series: | PLoS Biology |
| Online Access: | http://europepmc.org/articles/PMC6110572?pdf=render |
| _version_ | 1819149869836664832 |
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| author | Bryce A Mendelsohn Neal K Bennett Maxwell A Darch Katharine Yu Mai K Nguyen Daniela Pucciarelli Maxine Nelson Max A Horlbeck Luke A Gilbert William Hyun Martin Kampmann Jean L Nakamura Ken Nakamura |
| author_facet | Bryce A Mendelsohn Neal K Bennett Maxwell A Darch Katharine Yu Mai K Nguyen Daniela Pucciarelli Maxine Nelson Max A Horlbeck Luke A Gilbert William Hyun Martin Kampmann Jean L Nakamura Ken Nakamura |
| author_sort | Bryce A Mendelsohn |
| collection | DOAJ |
| description | Insufficient or dysregulated energy metabolism may underlie diverse inherited and degenerative diseases, cancer, and even aging itself. ATP is the central energy carrier in cells, but critical pathways for regulating ATP levels are not systematically understood. We combined a pooled clustered regularly interspaced short palindromic repeats interference (CRISPRi) library enriched for mitochondrial genes, a fluorescent biosensor, and fluorescence-activated cell sorting (FACS) in a high-throughput genetic screen to assay ATP concentrations in live human cells. We identified genes not known to be involved in energy metabolism. Most mitochondrial ribosomal proteins are essential in maintaining ATP levels under respiratory conditions, and impaired respiration predicts poor growth. We also identified genes for which coenzyme Q10 (CoQ10) supplementation rescued ATP deficits caused by knockdown. These included CoQ10 biosynthetic genes associated with human disease and a subset of genes not linked to CoQ10 biosynthesis, indicating that increasing CoQ10 can preserve ATP in specific genetic contexts. This screening paradigm reveals mechanisms of metabolic control and genetic defects responsive to energy-based therapies. |
| first_indexed | 2024-12-22T14:08:28Z |
| format | Article |
| id | doaj.art-e811f8fca40f46dda21b805657c1f66e |
| institution | Directory Open Access Journal |
| issn | 1544-9173 1545-7885 |
| language | English |
| last_indexed | 2024-12-22T14:08:28Z |
| publishDate | 2018-08-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Biology |
| spelling | doaj.art-e811f8fca40f46dda21b805657c1f66e2022-12-21T18:23:16ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852018-08-01168e200462410.1371/journal.pbio.2004624A high-throughput screen of real-time ATP levels in individual cells reveals mechanisms of energy failure.Bryce A MendelsohnNeal K BennettMaxwell A DarchKatharine YuMai K NguyenDaniela PucciarelliMaxine NelsonMax A HorlbeckLuke A GilbertWilliam HyunMartin KampmannJean L NakamuraKen NakamuraInsufficient or dysregulated energy metabolism may underlie diverse inherited and degenerative diseases, cancer, and even aging itself. ATP is the central energy carrier in cells, but critical pathways for regulating ATP levels are not systematically understood. We combined a pooled clustered regularly interspaced short palindromic repeats interference (CRISPRi) library enriched for mitochondrial genes, a fluorescent biosensor, and fluorescence-activated cell sorting (FACS) in a high-throughput genetic screen to assay ATP concentrations in live human cells. We identified genes not known to be involved in energy metabolism. Most mitochondrial ribosomal proteins are essential in maintaining ATP levels under respiratory conditions, and impaired respiration predicts poor growth. We also identified genes for which coenzyme Q10 (CoQ10) supplementation rescued ATP deficits caused by knockdown. These included CoQ10 biosynthetic genes associated with human disease and a subset of genes not linked to CoQ10 biosynthesis, indicating that increasing CoQ10 can preserve ATP in specific genetic contexts. This screening paradigm reveals mechanisms of metabolic control and genetic defects responsive to energy-based therapies.http://europepmc.org/articles/PMC6110572?pdf=render |
| spellingShingle | Bryce A Mendelsohn Neal K Bennett Maxwell A Darch Katharine Yu Mai K Nguyen Daniela Pucciarelli Maxine Nelson Max A Horlbeck Luke A Gilbert William Hyun Martin Kampmann Jean L Nakamura Ken Nakamura A high-throughput screen of real-time ATP levels in individual cells reveals mechanisms of energy failure. PLoS Biology |
| title | A high-throughput screen of real-time ATP levels in individual cells reveals mechanisms of energy failure. |
| title_full | A high-throughput screen of real-time ATP levels in individual cells reveals mechanisms of energy failure. |
| title_fullStr | A high-throughput screen of real-time ATP levels in individual cells reveals mechanisms of energy failure. |
| title_full_unstemmed | A high-throughput screen of real-time ATP levels in individual cells reveals mechanisms of energy failure. |
| title_short | A high-throughput screen of real-time ATP levels in individual cells reveals mechanisms of energy failure. |
| title_sort | high throughput screen of real time atp levels in individual cells reveals mechanisms of energy failure |
| url | http://europepmc.org/articles/PMC6110572?pdf=render |
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