Abstract
To explore possible relationships between mitochondrial DNA (mtDNA) polymorphism and the expression levels of stress-responder nuclear genes we assembled five cybrid cell lines by repopulating 143B.TK− cells, depleted of their own mtDNA (Rho0 cells), with foreign mitochondria with different mtDNA sequences (lines H, J, T, U, X). We evaluated, at both basal and under heat stress conditions, gene expression (mRNA) and intra-mitochondrial protein levels of HSP60 and HSP75, two key components in cellular stress response. At basal conditions, the levels of HSP60 and HSP75 mRNA were lower in one cybrid (H) than in the others (p = 0.005 and p = 0.001, respectively). Under stress conditions, the H line over-expressed both genes, so that the inter-cybrid difference was abolished. Moreover, the HSP60 intra-mitochondrial protein levels differed among the cybrid lines (p = 0.001), with levels higher in H than in the other cybrid lines. On the whole, our results provide further experimental evidence that mtDNA variability influences the cell response to stressful conditions by modulating components involved in this response. Sentence summary of the article: the results reported in the present study provide important experimental evidence that in human cells mtDNA variability is able to influence the cellular response to heat stress by modulating both the transcription of genes involved in this response and their intra-mitochondrial protein levels.
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Abbreviations
- ROS:
-
reactive oxygen species
- RFLP:
-
restriction fragment length polymorphism
- RT-PCR:
-
reverse-transcriptase-polymerase chain reaction
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Acknowledgements
This work was partly supported by Fondo Sociale Europeo—FSE (PhD course in Molecular Biopathology, University of Calabria, Italy) and by University Grants for Scientific Research (ex 60%).
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D. Bellizzi and D. Taverna equally contributed to this study.
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RFLP analysis of common variability in parental and cybrid cell lines. +/− indicate the presence/absence of the restriction site with reference to the MITOMAP sequence (www.mitomap.org). (DOC 575 KB)
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Bellizzi, D., Taverna, D., D’Aquila, P. et al. Mitochondrial DNA variability modulates mRNA and intra-mitochondrial protein levels of HSP60 and HSP75: experimental evidence from cybrid lines. Cell Stress and Chaperones 14, 265–271 (2009). https://doi.org/10.1007/s12192-008-0081-x
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DOI: https://doi.org/10.1007/s12192-008-0081-x