Heavy Water Reduces GFP Expression in Prokaryotic Cell-Free Assays at the Translation Level While Stimulating Its Transcription

The in vitro proliferation of prokaryotic and eukaryotic cells is remarkably hampered in the presence of heavy water (D2O). Impairment of gene expression at the transcription or translation level can be the base for this effect. However, insights into the underlying mechanisms are lacking. Here, we...

Full description

Saved in:
Bibliographic Details
Published in:BioMed research international 2013-01, Vol.2013 (2013), p.1-9
Main Authors: Bayerl, Thomas M., Opitz, Madeleine, Stögbauer, Tobias, Hohlefelder, Luisa S., Rädler, Joachim O.
Format: Article
Language:English
Subjects:
Animals
Cell division
Cell-Free System
Data analysis
Deoxyribonucleic acid
Deuterium Oxide - pharmacology
DNA
E coli
Enzymes
Escherichia coli - drug effects
Gene expression
Gene Expression Regulation - drug effects
Genetic transcription
Green fluorescent protein
Green Fluorescent Proteins - biosynthesis
Heavy water
Protein Biosynthesis - drug effects
Protein expression
Proteins
RNA polymerase
RNA, Messenger - genetics
Software
Transcription, Genetic - drug effects
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The in vitro proliferation of prokaryotic and eukaryotic cells is remarkably hampered in the presence of heavy water (D2O). Impairment of gene expression at the transcription or translation level can be the base for this effect. However, insights into the underlying mechanisms are lacking. Here, we employ a cell-free expression system for the quantitative analysis of the effect of increasing percentages of D2O on the kinetics of in-vitro GFP expression. Experiments are designed to discriminate the rates of transcription, translation, and protein folding using pDNA and mRNA vectors, respectively. We find that D2O significantly stimulates GFP expression at the transcription level but acts as a suppressor at translation and maturation (folding) in a linear dose-dependent manner. At a D2O concentration of 60%, the GFP expression rate was reduced to 40% of an undisturbed sample. We observed a similar inhibition of GFP expression by D2O in a recombinant Escherichia coli strain, although the inhibitory effect is less pronounced. These results demonstrate the suitability of cell-free systems for quantifying the impact of heavy water on gene expression and establish a platform to further assess the potential therapeutic use of heavy water as antiproliferative agent.
ISSN:2314-6133
2314-6141
DOI:10.1155/2013/592745