Apoptosis

, Volume 10, Issue 6, pp 1469–1482

Differential gene expression during apoptosis induced by a serum factor: Role of mitochondrial F0-F1 ATP synthase complex

Article

Abstract

The number of genes that are up regulated or down regulated during apoptosis is large and still increasing. In an attempt to characterize differential gene expression during serum factor induced apoptosis in AK-5 cells (a rat histiocytoma), we found subunit 6 and subunit 8 of the transmembrane proton channel and subunit alpha of the catalytic core of the mitochondrial F0-F1 ATP synthase complex to be up regulated during apoptosis. The increase in the expression levels of these subunits was concomitant with a transient increase in the intracellular ATP levels, suggesting that the increase in cellular ATP content is a result of the increase in the expression of ATP synthase subunits' gene and de novo protein synthesis. Depleting the cellular ATP levels with oligomycin inhibited apoptosis significantly, pointing to the requirement of ATP during apoptosis. Caspase 1 and caspase 3 activity and the loss of mitochondrial membrane potential were also inhibited by oligomycin during apoptosis in these cells, suggesting that the oligomycin induced inhibition of apoptosis could be due to inhibition of caspase activity and inhibition of mitochondrial depolarization. However, cytochrome C release during apoptosis was found to be completely independent of intracellular ATP content. Besides the ATP synthase complex genes, other mitochondrial genes like cytochrome C oxidase subunit II and III also showed elevated levels of expression during apoptosis. This kind of a mitochondrial gene expression profile suggests that in AK-5 cells, these genes are upregulated in a time-linked manner to ensure sufficient intracellular ATP levels and an efficient functioning of the mitochondrial respiratory chain for successful completion of the apoptotic pathway.

Keywords

apoptosis ATP synthase DD-PCR gene expression intracellular ATP mitochondria 

Abbreviations

Annexin V-FITC

fluorescein conjugated annexin V

TBST

Tris Borate Sodium Tween

SF

Serum factor

MRC

mitochondrial respiratory chain

Dex

Dexamethasone

DD-PCR

Differential Display PCR

Act D

Actinomycin D

CHX

Cycloheximide

2-DG

2-deoxyglucose

DEVD-AMC

Asp-Glu-Val-Asp-7-Amino-4-methylcoumarin

YVAD-AMC

Tyr-Val-Ala-Asp-AMC

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Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  1. 1.Centre for Cellular & Molecular BiologyHyderabadIndia

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