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  • Effects of metal ions and hydrogen peroxide on the phenotype of yeast hom6Δ mutant.

Effects of metal ions and hydrogen peroxide on the phenotype of yeast hom6Δ mutant.

Letters in applied microbiology (2014-10-07)
N M Tun, B R Lennon, P J O'Doherty, A J Johnson, G Petersingham, T D Bailey, C Kersaitis, M J Wu
ABSTRACT

HOM6 is a major gene in the aspartate pathway which leads to biosynthesis of threonine and methionine. The phenotypes of the gene deletion mutant (hom6∆) in a variety of cultural conditions have previously provided meaningful insights into the biological roles of HOM6 and its upstream intermediate metabolites. Here, we conducted a survey on a spectrum of metal ions for their effect on the aspartate pathway and broader sulphur metabolism. We show that manganese (Mn(2+) ) promoted the growth of hom6∆ under both anaerobic and aerobic conditions. Unexpectedly, 4 mmol l(-1) hydrogen peroxide (H2 O2 ), a dose normally causing temporary cell growth arrest, enhanced the growth of hom6∆ under the anaerobic condition only, while it had no effect on the wild type strain BY4743. We propose that Mn(2+) and H2 O2 promote the growth of hom6∆ by reducing the accumulation of the toxic intermediate metabolite-aspartate β-semialdehyde, via directing the aspartate pathway to the central sugar metabolism-tricarboxylic acid cycle. This study focuses on the yeast strain which lacks homoserine dehydrogenase encoded by HOM6 gene in aspartate metabolism. The HOM6-deletion mutant (hom6Δ) was analysed in the context of varying environmental parameters such as metal ions and oxidants, under anaerobic and aerobic conditions. We demonstrated that both manganese and hydrogen peroxide can promote the growth of hom6Δ, with the latter exerting such effect only under anaerobic condition. The findings are relevant to the research areas of ageing and anti-fungal drug development. It highlights the importance of interactions between gene expression and environmental factors as well as culture conditions.

MATERIALS
Product Number
Brand
Product Description

Supelco
L-Aspartic acid, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
L-Aspartic acid, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
Manganese, chips, thickness <2.0 mm, 99%
Sigma-Aldrich
Manganese, powder, −325 mesh, ≥99% trace metals basis
Sigma-Aldrich
Manganese, powder, ≥99.9% trace metals basis
Sigma-Aldrich
Hydrogen peroxide solution, 34.5-36.5%
SAFC
L-Aspartic acid
Sigma-Aldrich
DL-Aspartic acid, ≥99% (TLC)
Aspartic acid, European Pharmacopoeia (EP) Reference Standard
Supelco
L-Aspartic acid, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
L-Aspartic acid, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
L-Aspartic acid, BioUltra, ≥99.5% (T)
Sigma-Aldrich
L-Aspartic acid, BioXtra, ≥99% (HPLC)
Sigma-Aldrich
Hydrogen peroxide solution, contains ~200 ppm acetanilide as stabilizer, 3 wt. % in H2O
Millipore
Hydrogen peroxide solution, 3%, suitable for microbiology
Manganese, foil, 25x25mm, thickness 2mm, hot-pressed, 99.95%
Manganese, rod, 50mm, diameter 6.35mm, cast, 99.5%
Manganese, foil, not light tested, 25x25mm, thickness 0.02mm, permanent polyester support, 98.7%
Manganese, foil, not light tested, 25x25mm, thickness 0.015mm, permanent polyester support, 98.7%
Manganese, foil, not light tested, 50x50mm, thickness 0.02mm, permanent polyester support, 98.7%
Manganese, foil, not light tested, 25x25mm, thickness 0.007mm, permanent polyester support, 98.7%
Manganese, foil, not light tested, 25x25mm, thickness 0.009mm, permanent polyester support, 98.7%
Manganese, foil, not light tested, 50x50mm, thickness 0.015mm, permanent polyester support, 98.7%
Manganese, foil, not light tested, 25x25mm, thickness 0.008mm, permanent polyester support, 98.7%
Manganese, rod, 10mm, diameter 4.0mm, cast, 99.5%
Manganese, foil, not light tested, 25x25mm, thickness 0.0125mm, permanent polyester support, 98.7%
Manganese, foil, not light tested, 25x25mm, thickness 0.0025mm, permanent polyester support, 98.7%
Manganese, foil, not light tested, 25x25mm, thickness 0.01mm, permanent polyester support, 98.7%
Manganese, foil, not light tested, 25x25mm, thickness 0.003mm, permanent polyester support, 98.7%
Manganese, foil, not light tested, 25x25mm, thickness 0.05mm, permanent polyester support, 98.7%