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Green Fluorescent Protein Alters the Transcriptional Regulation of Human Mitochondrial Genes After Gamma Irradiation

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Abstract

Green fluorescent proteins (GFP), extensively used as reporters in biological and imaging studies, are assumed to be mostly biologically inert. Here, we test the assumption in regard to the transcriptional regulation of 18 mitochondrially encoded genes in GFP expressing human T-cell line (JURKAT cells) exposed to gamma radiation. Using quantitative polymerase chain reaction, we demonstrate that wild type and GFP expressing JURKAT cells have different baseline mitochondrial transcript expression (10 out of the 18 tested genes) and after a single dose of radiation (100 Gy) show a significantly different transcriptional regulation of their mitochondrial genes. While in wild type cells, ten of the tested genes are up-regulated in response to radiation exposure, GFP expressing cells show less transcriptional regulation with a small down-regulation in five genes. Our results indicate that the presence of GFP in the cytoplasm can alter the cellular response to ionizing radiation.

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Abbreviations

GFP:

Green fluorescent protein

EmGFP:

Emerald green fluorescent protein

FBS:

Fetal bovine serum

DMEM:

Dulbecco’s modified eagle medium

qPCR:

Quantitative polymerase chain reaction

DNA:

Deoxyribonucleic acid

RNA:

Ribonucleic acid

mRNA:

Messenger ribonucleic acid

tRNA:

Transfer ribonucleic acid

rRNA:

Ribosomal ribonucleic acid

SD:

Standard deviation

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Acknowledgments

Special thanks should be given to Dr Guo Jun Lin for providing the fluorescent microscopic image. ANSTO’s External Radiation Group, for their technical assistance in performing the irradiation experiments and provision of technical details for the “Gamma irradiation” section. Dr Anya Salih, for discussing the manuscript.

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Authors and Affiliations

  1. Australian Nuclear Science and Technology Organisation, Lucas Heights, Sydney, New South Wales, 2234, Australia

    Winnie Wai-Ying Kam, Ryan Middleton, Vanessa Lake & Richard B. Banati

  2. National Imaging Facility at Brain and Mind Research Institute (BMRI), University of Sydney, Camperdown, Sydney, New South Wales, 2050, Australia

    Richard B. Banati

  3. Medical Radiation Sciences, Faculty of Health Sciences, University of Sydney, Cumberland, Sydney, New South Wales, 2141, Australia

    Winnie Wai-Ying Kam & Richard B. Banati

Authors
  1. Winnie Wai-Ying Kam
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  2. Ryan Middleton
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Correspondence to Winnie Wai-Ying Kam.

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Kam, W.WY., Middleton, R., Lake, V. et al. Green Fluorescent Protein Alters the Transcriptional Regulation of Human Mitochondrial Genes After Gamma Irradiation. J Fluoresc 23, 613–619 (2013). https://doi.org/10.1007/s10895-013-1206-x

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  • DOI: https://doi.org/10.1007/s10895-013-1206-x

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