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Susceptibility of HIV-1-TAT transfected cells to undergo apoptosis. Biochemical mechanisms

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Abstract

The effects of HIV-1 Tat protein on mitochondria membrane permeability and apoptosis were analysed in lymphoid cells. In this report we show that stable-transfected HIV-Tat cells are primed to undergo apoptosis upon serum withdrawal. This effect was observed in both the Jhan T cell line and the K562 cells, the latter expressing the bcr-abl chimeric gene, which confers resistance to apoptosis induced by different stimuli. Using a cytofluorimetric approach we have determined that serum withdrawal induces a disruption of the transmembrane mitochondrial potential (Δψm) followed by an increase of reactive oxygen species (ROS) and the subsequent DNA nuclear loss in K562-Tat cells but not in the K562-pcDNA cell line. These pre-apoptotic events were associated with the cleavage of the caspase-3, while the expression of Bcl-2, Bcl-XL and Bax proteins was not affected by the presence of Tat. Regardless of the steady state of the Bax protein, we found that in both K562 and K562-Tat cells, this protein is located in the nucleus, but after serum withdrawal its localization was mainly in the cytoplasm. The activity of caspase-3 detected in K562-Tat cells after serum withdrawal paralleled with the mitochondria permeability transition. Nevertheless, in Jhan-Tat cells the inhibition of this caspase with the specific inhibitor, z-DEVD-cmk, did not affect the disruption of the mitochondria potential induced by serum withdrawal. Interestingly, we found that HIV-Tat protein accumulates at the mitochondria in the K562-Tat cells cultured under low serum conditions, and this mitochondrial localization correlated with the Δψm disruption detected in these cells. In addition, HIV-1 Tat protein synergies with protoporphyrin IX (PPIX), a ligand of the mitochondrial benzodiazepine receptor, in the induction of apoptosis in both Jhan and K562 cells. Thus, HIV-1 Tat protein may induce apoptosis by a mechanism that involves mitochondrial PT and may contribute to the lymphocyte depletion seen in AIDS patients.

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Abbreviations

Δψm:

mitochondrial transmembrane potential

DiOC6(3):

3,3′dihexyloxacarbocyanine iodide

HE:

hydroethidine

PI:

propidium iodide

PT:

permeability transition

PPIX:

Protoporphyrin IX

ROS:

reactive oxygen species

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Acknowledgements

This work was supported by CICYT grant to SAF98-0046 to E Muñoz. The authors wish to thank Ms Carmen Cabrero-Doncel for assistance with the manuscript.

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Authors and Affiliations

  1. Departamento de Fisiología e Inmunología, Universidad de Córdoba, Facultad de Medicina, Avda, de Menéndez Pidal s/n, 14071, Córdoba, Spain

    Antonio Macho, Marco A Calzado, Luis Jiménez-Reina & Eduardo Muñoz

  2. Departamento de Bioquímica y Biología Molecular, Universidad de Cantabria, Spain

    Eva Ceballos & Javier León

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  1. Antonio Macho
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Macho, A., Calzado, M., Jiménez-Reina, L. et al. Susceptibility of HIV-1-TAT transfected cells to undergo apoptosis. Biochemical mechanisms. Oncogene 18, 7543–7551 (1999). https://doi.org/10.1038/sj.onc.1203095

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