Journal of Fluorescence

, Volume 23, Issue 4, pp 613–619

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

  • Winnie Wai-Ying Kam
  • Ryan Middleton
  • Vanessa Lake
  • Richard B. Banati
SHORT COMMUNICATION

DOI: 10.1007/s10895-013-1206-x

Cite this article as:
Kam, W.WY., Middleton, R., Lake, V. et al. J Fluoresc (2013) 23: 613. doi:10.1007/s10895-013-1206-x

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.

Keywords

Green fluorescent protein JURKAT cells Mitochondria Gene expression Aequorea victoria Gamma radiation 

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

Copyright information

© © Crown Copyright as represented by: Australian Nuclear Science and Technology Organisation 2013

Authors and Affiliations

  • Winnie Wai-Ying Kam
    • 1
    • 3
  • Ryan Middleton
    • 1
  • Vanessa Lake
    • 1
  • Richard B. Banati
    • 1
    • 2
    • 3
  1. 1.Australian Nuclear Science and Technology OrganisationSydneyAustralia
  2. 2.National Imaging Facility at Brain and Mind Research Institute (BMRI)University of SydneySydneyAustralia
  3. 3.Medical Radiation Sciences, Faculty of Health SciencesUniversity of SydneySydneyAustralia

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