Comparative analysis of nuclear proteomes in mitochondrial DNA-depleted A549 cells and their parental cells
10.3867/j.issn.1000-3002.2012.04.003
- VernacularTitle:线粒体DNA缺失A549细胞与其母本细胞核蛋白质组差异分析
- Author:
Peng ZHAO
;
Zhaohui ZHANG
;
Weijian ZHONG
;
Xianping YING
;
Zhun YUANN
;
Biyun YAO
;
Juanling FU
;
Zongcan ZHOU
- Publication Type:Journal Article
- Keywords:
nuclear proteomics;
genes,p53;
membrane potential,mitochondrial
- From:
Chinese Journal of Pharmacology and Toxicology
2012;26(4):482-488
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To investigate the nuclear proteomes in mitochondrial DNA (mtDNA)-depleted A549 cells (Rho0 cells) and their parental cells (Rho+ cells),and to learn more about the nuclear responses to mitochondrial dysfunction.METHODS The nuclear proteomes of Rho and Rho + cells were characterized by two dimensional electrophoresis (2-DE) and SELDI-TOF ProteinChip technologies,the differentially expressed protein- spots were identified by MALDI-TOF mass spectrum (MS),the nucleophosmin and P53 expression were detected by Western blotting assay,and the mitochondrial memhrane potential (MMP) was measured by the laser scanning confocal microscope.RESULTS 2-DE results showed 11 protein-spots were down-regulated and 21 protein-spots were up-regulated in Rho0 cell nuclei.SELDI-TOF MS analysis with NP20 ProteinChips revealed 4 protein-peaks decreased in Rho0 cell nuclei.One down-regulated protein-spot was identified as elF-6,and 4 up-regulated proteinspots were identified as nucleophosmin,SFRS1,SFRS3 and hnRNP G,respectively.The increased expression of nucleophosmin in Rho0 cells was verified by Western blotting.Based on the clues from proteomic analysis,P53 expression in Rho0 cells was higher than in Rho + cells,and MMP was consistent in Rho + and Rho0 cells.CONCLUSION mtDNA-depletion induces nuclear proteome alteration.Rho0 cells can be used as a model to study the crosstalk between mitochondrion and nucleus.
