Application of the comprehensive set of heterozygous yeast deletion mutants to elucidate the molecular basis of cellular chromium toxicity

Sara Holland¹ , Emma Lodwig¹ , Theodora Sideri¹ , Tom Reader¹ , Ian Clarke² , Konstantinos Gkargkas³ , David C Hoyle² , Daniela Delneri⁴ , Stephen G Oliver³ and Simon V Avery¹

¹School of Biology, Institute of Genetics, The University of Nottingham, University Park, Nottingham NG7 2RD, UK

²North West Institute for Bio-Health Informatics, The University of Manchester, ISBE, School of Medicine, Oxford Road, Manchester M13 9PT, UK

³Department of Biochemistry, University of Cambridge, Sanger Building, Tennis Court Road, Cambridge CB2 1GA, UK

⁴Faculty of Life Sciences, The University of Manchester, Oxford Road, Manchester M13 9PT, UK

author email corresponding author email

Genome Biology 2007, 8:R268doi:10.1186/gb-2007-8-12-r268


Published:	18 December 2007

Subject areas: Genetics, Genome studies, Molecular biology, Physiology

Abstract

Background

The serious biological consequences of metal toxicity are well documented, but the key modes of action of most metals are unknown. To help unravel molecular mechanisms underlying the action of chromium, a metal of major toxicological importance, we grew over 6,000 heterozygous yeast mutants in competition in the presence of chromium. Microarray-based screens of these heterozygotes are truly genome-wide as they include both essential and non-essential genes.

Results

The screening data indicated that proteasomal (protein degradation) activity is crucial for cellular chromium (Cr) resistance. Further investigations showed that Cr causes the accumulation of insoluble and toxic protein aggregates, which predominantly arise from proteins synthesised during Cr exposure. A protein-synthesis defect provoked by Cr was identified as mRNA mistranslation, which was oxygen-dependent. Moreover, Cr exhibited synergistic toxicity with a ribosome-targeting drug (paromomycin) that is known to act via mistranslation, while manipulation of translational accuracy modulated Cr toxicity.

Conclusion

The datasets from the heterozygote screen represent an important public resource that may be exploited to discover the toxic mechanisms of chromium. That potential was validated here with the demonstration that mRNA mistranslation is a primary cause of cellular Cr toxicity.