Format

Send to

Choose Destination
Drug Discov Ther. 2019;13(6):306-313. doi: 10.5582/ddt.2019.01080.

Gene disruption of ribosomal protein L5 (RPL5) decreased the sensitivity of CHO-K1 cells to uncoupler carbonylcyanide-3-chlorophenylhydrazone.

Author information

1
Meiji Pharmaceutical University, Tokyo, Japan.
2
Ritsumeikan-Global Innovation Research Organization, Ritsumeikan University, Kyoto, Japan.
3
Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
4
School of Pharmacy, Iwate Medical University, Iwate, Japan.
5
Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan.
6
Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences, Niigata, Japan.

Abstract

Protonophoric uncoupler carbonylcyanide-3-chlorophenylhydrazone (CCCP) decreases the proton motive force (ΔP) of the mitochondrial inner membrane and results in inhibition of oxidative phosphorylation. In this study, a CCCP-resistant clone was isolated from a random gene trap insertional mutant library of Chinese hamster ovary (CHO)-K1 cells which was constructed by infecting a retrovirus vector, ROSAβgeo. Although we expected the isolation of the mutants defective in nuclear genes responsible for mitochondrial functions, the disrupted gene of the isolated mutant that we named R1 cells was identified as one of the alleles for ribosomal protein 5 of large subunit (RPL5). The R1 cells express as much as 80% RPL5 protein compared with the parental CHO-K1 cells, possibly due to enhanced transcription from a remaining wild-type RPL5 allele in R1 cells. Furthermore, the protein amount is not decreased by CCCP in R1 cells, in contrast to its clear reduction by CCCP in parental cells. Since mutations of RPL5 and other ribosomal proteins are responsible for the ribosomopathies and cancer, the present mutant may be a useful cellular model of such human diseases from a viewpoint of energy metabolism as well as a tool for the study of ribosome biogenesis and extra-ribosomal function of the RPL5 protein.

KEYWORDS:

Diamond-Blackfan anemia; Ribosomopathies; mitochondria; random gene trap mutagenesis; uncoupler

PMID:
31956228
DOI:
10.5582/ddt.2019.01080
Free full text

Supplemental Content

Full text links

Icon for J-STAGE, Japan Science and Technology Information Aggregator, Electronic
Loading ...
Support Center