Mathematical modelling of arsenic transport, distribution and detoxification processes in yeast.

Arsenic has a dual role as causative and curative agent of human disease. Therefore, there is considerable interest in elucidating arsenic toxicity and detoxification mechanisms. By an ensemble modelling approach, we identified a best parsimonious mathematical model which recapitulates and predicts intracellular arsenic dynamics for different conditions and mutants, thereby providing novel insights into arsenic toxicity and detoxification mechanisms in yeast, which could partly be confirmed experimentally by dedicated experiments. Specifically, our analyses suggest that: (i) arsenic is mainly protein-bound during short-term (acute) exposure, whereas glutathione-conjugated arsenic dominates during long-term (chronic) exposure, (ii) arsenic is not stably retained, but can leave the vacuole via an export mechanism, and (iii) Fps1 is controlled by Hog1-dependent and Hog1-independent mechanisms during arsenite stress. Our results challenge glutathione depletion as a key mechanism for arsenic toxicity and instead suggest that (iv) increased glutathione biosynthesis protects the proteome against the damaging effects of arsenic and that (v) widespread protein inactivation contributes to the toxicity of this metalloid. Our work in yeast may prove useful to elucidate similar mechanisms in higher eukaryotes and have implications for the use of arsenic in medical therapy.

The MAPK Hog1p modulates Fps1p-dependent arsenite uptake and tolerance in yeast.

Arsenic is widely distributed in nature and all organisms possess regulatory mechanisms to evade toxicity and acquire tolerance. Yet, little is known about arsenic sensing and signaling mechanisms or about their impact on tolerance and detoxification systems. Here, we describe a novel role of the S. cerevisiae mitogen-activated protein kinase Hog1p in protecting cells during exposure to arsenite and the related metalloid antimonite. Cells impaired in Hog1p function are metalloid hypersensitive, whereas cells with elevated Hog1p activity display improved tolerance. Hog1p is phosphorylated in response to arsenite and this phosphorylation requires Ssk1p and Pbs2p. Arsenite-activated Hog1p remains primarily cytoplasmic and does not mediate a major transcriptional response. Instead, hog1delta sensitivity is accompanied by elevated cellular arsenic levels and we demonstrate that increased arsenite influx is dependent on the aquaglyceroporin Fps1p. Fps1p is phosphorylated on threonine 231 in vivo and this phosphorylation critically affects Fps1p activity. Moreover, Hog1p is shown to affect Fps1p phosphorylation. Our data are the first to demonstrate Hog1p activation by metalloids and provides a mechanism by which this kinase contributes to tolerance acquisition. Understanding how arsenite/antimonite uptake and toxicity is modulated may prove of value for their use in medical therapy.

A subgroup of plant aquaporins facilitate the bi-directional diffusion of As(OH)3 and Sb(OH)3 across membranes.

BACKGROUND: Arsenic is a toxic and highly abundant metalloid that endangers human health through drinking water and the food chain. The most common forms of arsenic in the environment are arsenate (As(V)) and arsenite (As(III)). As(V) is a non-functional phosphate analog that enters the food chain via plant phosphate transporters. Inside cells, As(V) becomes reduced to As(III) for subsequent extrusion or compartmentation. Although much is known about As(III) transport and handling in microbes and mammals, the transport systems for As(III) have not yet been characterized in plants. RESULTS: Here we show that the Nodulin26-like Intrinsic Proteins (NIPs) AtNIP5;1 and AtNIP6;1 from Arabidopsis thaliana, OsNIP2;1 and OsNIP3;2 from Oryza sativa, and LjNIP5;1 and LjNIP6;1 from Lotus japonicus are bi-directional As(III) channels. Expression of these NIPs sensitized yeast cells to As(III) and antimonite (Sb(III)), and direct transport assays confirmed their ability to facilitate As(III) transport across cell membranes. On medium containing As(V), expression of the same NIPs improved yeast growth, probably due to increased As(III) efflux. Our data furthermore provide evidence that NIPs can discriminate between highly similar substrates and that they may have differential preferences in the direction of transport. A subgroup of As(III) permeable channels that group together in a phylogenetic tree required N-terminal truncation for functional expression in yeast. CONCLUSION: This is the first molecular identification of plant As(III) transport systems and we propose that metalloid transport through NIPs is a conserved and ancient feature. Our observations are potentially of great importance for improved remediation and tolerance of plants, and may provide a key to the development of low arsenic crops for food production.

Assessment of inorganic content of PM(2.5) particles sampled in a rural area north-east of Hanoi, Vietnam.

Atmospheric aerosols from seven rural sites in northern Vietnam, east of Hanoi, were sampled and analyzed. The aim of the study was to evaluate trace elemental and black carbon (BC) concentrations in fine particles (PM(2.5)) and to investigate the influence of the Pha Lai power plant and other pollution sources on regional air quality. Seven measurement stations were set up at selected rural sites and a campaign consisting of 12 two-day measurement periods was conducted from the end of May until the end of October 2000. At each location a pair of samplers was installed consisting of a cyclone loaded with Teflon filters and a modified Millipore air monitoring cartridge loaded with glass fibre filters. The obtained samples were analyzed for trace elements by Energy Dispersive X-ray Fluorescence (EDXRF), while a black smoke detector was used for BC analysis. Seventeen trace elements were analyzed; Br, Ca, Cl, Cr, Cu, Fe, K, Mn, Ni, Pb, Rb, S, Se, Sr, Ti, V and Zn and their concentrations evaluated. The results showed that BC, Ca, Cl, Fe, K and S dominated in the sampled atmospheric aerosols. The measured concentrations of the potentially hazardous trace elements Cr, Mn, Ni and Pb were all below the limits defined by Vietnamese standards of ambient air quality. Statistical evaluations indicated that coal and heavy fuel oil combustion were major sources of atmospheric pollutants in the area and that biomass burning and road transport had a marked influence on regional air quality. It was concluded that the Pha Lai power plant was the major source of coal combustion emissions. Trace element emissions originating from river transportation were suggested as another major source of atmospheric pollutants. The results indicate that the elemental concentrations in PM(2.5) are strongly influenced by seasonal variations. Further measurements are required to identify the impact of long-range transported continental air masses on the air quality of the investigated rural area.

Elemental composition of tropospheric aerosols in Hanoi, Vietnam and Nairobi, Kenya.

Air pollution problems in major cities within the developing countries need to be studied. There are scanty measurements from the developing countries on airborne particles despite their adverse implications to human health, visibility and climate. One of the major sources of anthropogenic air pollution is energy production. Energy demand is bound to increase as population increases, especially in major cities of the world. Fine particles, particles with aerodynamic diameter < or = 2.5 microm, are mainly anthropogenic and these particles were collected in the capital cities of Vietnam and Kenya. A cyclone airborne particle collector was used to sample in Hanoi during the months of May to October 2000 and a dichotomous virtual impactor in Nairobi in February 2000. The samples were analysed for elemental content by an energy dispersive X-ray fluorescence (EDXRF) spectrometer. S, Cl, K and Fe exceeded atmospheric concentrations of 100 ng m(-3) at both cities. Atmospheric elemental concentrations in both Hanoi and Nairobi were orders of magnitude higher than their respective rural towns. Traffic, biomass and waste burning emissions were implicated as the main sources of air pollution in Nairobi, while coal combustion and road transport were the major sources in Hanoi. Regional air pollution had a major impact over Hanoi, whereas an influence of that kind was not identified in Nairobi. Pb and other toxic elements had concentration levels below WHO guideline, however, the two cities are threatened by future high levels of air pollution due to the high rate of population growth. Long-term measurements are required in both areas to evaluate if the alarming situation is deteriorating.