A genome-wide shRNA screen for new OxPhos related genes

The mitochondrial oxidative phosphorylation (OxPhos) system produces most of the ATP required by the cell. The structural proteins of the OxPhos holoenzymes are well known, but important aspects of their biogenesis and regulation remain to be uncovered and a significant fraction of mitochondrial pro...

Full description

Saved in:
Bibliographic Details
Published in:Mitochondrion 2011-05, Vol.11 (3), p.467-475
Main Authors: Bayona-Bafaluy, María Pilar, Sánchez-Cabo, Fátima, Fernández-Silva, Patricio, Pérez-Martos, Acisclo, Enríquez, José Antonio
Format: Article
Language:English
Subjects:
Animals
Cell Line
Electron Transport Chain Complex Proteins - genetics
Electron Transport Chain Complex Proteins - metabolism
Fibroblasts - metabolism
Genetic Vectors
High-Throughput Screening Assays - methods
Lentivirus - genetics
Mice
Mitochondria - enzymology
Mitochondria - metabolism
Oxidative Phosphorylation
Oxidoreductases - antagonists & inhibitors
Oxidoreductases - genetics
RNA Interference
RNA, Small Interfering - genetics
RNA, Small Interfering - metabolism
Transduction, Genetic
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The mitochondrial oxidative phosphorylation (OxPhos) system produces most of the ATP required by the cell. The structural proteins of the OxPhos holoenzymes are well known, but important aspects of their biogenesis and regulation remain to be uncovered and a significant fraction of mitochondrial proteins have yet to be identified. We have used a high throughput, genome-wide RNA interference (RNAi) approach to identify new OxPhos-related genes. We transduced a mouse fibroblast cell line with a lentiviral-based shRNA-library, and screened the cell population for growth impairment in galactose-based medium, which requires an intact OxPhos system. Candidate genes were ranked according to their co-expression with known genes encoding OxPhos mitochondria-located proteins. For the top ranking candidates the cellular process in which they are involved was evaluated. Our results show that the use of genome-wide RNAi together with screening for deficient growth in galactose medium is a suitable approach to identifying OxPhos-related and cellular energy metabolism-related genes. Interestingly also ubiquitin-proteasome related genes were selected.
ISSN:1567-7249
1872-8278
DOI:10.1016/j.mito.2011.01.007