No description available.

Copper and iron are trace elements that form an indispensable part of many proteins and are crucial for the well-being of all cells. At the same time, the intracellular levels of these metals require careful regulation, as excess or deficiency may be lethal. P1B-ATPases are key players in metal homeostasis. They belong to the superfamily of P-type ATPases, transmembrane proteins present in virtually all life forms, are responsible for solute translocation across biological membranes. The goal of this thesis is to improve our understanding of the structural and functional roles of P1B-ATPases in metal homeostasis by focusing on the host-microbe interaction. The thesis first describes the importance of Cu+ distribution in the outcome of host-microbe interaction. Copper is an important element in host-microbe interactions, acting both as a catalyst in enzymes and as a potential toxin. Cu+-ATPases drive cytoplasmic Cu+ efflux and protect bacteria against metal overload. Many pathogenic and symbiotic bacteria contain multiple Cu+-ATPase genes within particular genetic environments, suggesting alternative roles for each resulting protein. This hypothesis was tested by characterizing five homologous Cu+-ATPases present in the symbiotic organism Sinorhizobium meliloti. Mutation of each gene led to different phenotypes and abnormal nodule development in the alfalfa host. Distinct responses were detected in free-living S. meliloti mutant strains exposed to metal and redox stresses. Differential gene expression was detected under Cu+, oxygen or nitrosative stress. These observations suggest that CopA1a maintains the cytoplasmic Cu+ quota and its expression is controlled by Cu+ levels. CopA1b is also regulated by Cu+ concentrations and is required during symbiosis for bacteroid maturation. CopA2-like proteins, FixI1 and FixI2, are necessary for the assembly of two different cytochrome c oxidases at different stages of bacterial life. CopA3 is a phylogenetically distinct Cu+-ATPase that does not contribute to Cu+ tolerance. It is regulated by redox stress and required during symbiosis. We postulated a model where non-redundant homologous Cu+-ATPases, operating under distinct regulation, transport Cu+ to different target proteins. In its second part, the thesis describes the novel Fe2+-ATPases and their influence in the host-microbe interaction. Little is known about iron efflux transporters in bacterial systems. Recently, the participation of Bacillus subtilis PfeT, a P1B4-ATPase, in cytoplasmic Fe2+ efflux has been proposed. We report here the distinct roles of mycobacterial P1B4-ATPases in the homeostasis of Co2+ and Fe2+. Mutation of Mycobacterium smegmatis ctpJ affects the homeostasis of both ions. Alternatively, a M. tuberculosis ctpJ mutant is more sensitive to Co2+ than Fe2+, while mutation of the homologous M. tuberculosis ctpD leads to Fe2+ sensitivity but no alterations in Co2+ homeostasis. In vitro, the three enzymes are activated by both Fe2+ and Co2+ and bind one equivalent of either ion at their transport site. However, equilibrium binding affinities and activity kinetics show that M. tuberculosis CtpD has higher affinity for Fe2+ and twice the Fe2+ stimulated activity than the CtpJs. These parameters are accompanied by a lower activation by and affinity for Co2+. Analysis of Fe2+ and Co2+ binding to CtpD by X-ray spectroscopy shows that both ions are coordinated by 5-6 O/N atoms with similar geometry. Mutagenesis studies suggest the involvement of invariant Ser, His and Glu in metal coordination. Interestingly, replacement of Cys in the conserved CPS sequence at the metal binding pocket leads to a large reduction in Fe2+ but not Co2+ binding affinity. We propose that CtpJ ATPases participate in the control of steady state Fe2+ levels. CtpD, required for M. tuberculosis virulence, is a high affinity Fe2+ transporter involved in the rapid response to iron dyshomeostasis generated upon redox stress. These studies provide significant insights into the metal selectivity, regulation, transport kinetics and functional diversity of homologous P1B-ATPases in Cu+ and Fe2+ homeostasis. Moreover, these biochemical characterizations can be integrated with the structural-functional analysis to elucidate the complex metal distribution networks.

The boreal landscape is complex mosaic of vast forests, lakes and wetlands. Through the landscape flows a fine network of streams and rivers, carrying dissolved and suspended material from atmospheric deposition and weathering of soils and bedrock to downstream recipients. This thesis investigates the transport of U, Th and other metals in the boreal landscape by comparing a set of catchments with contrasting characteristics, ranging from 0.12-68 km2 in area. Using uranium (234U/238U) and oxygen isotopes (δ18O) it was demonstrated that catchment size has a strong impact on the hydrological pathways and on the mobilisation of uranium. Both tracers also displayed a consistent shift towards more superficial sources and more superficial flow pathways when going from winter baseflow conditions to the spring flood. Large spatiotemporal variability was observed with U fluxes ranging from 1.7 -30 g km-2 a-1. Using a wide set of hydrochemical parameters and landscape characteristics it was demonstrated that wetlands play a decisive role for the biogeochemical cycling of many metals. Comparing normalised fluxes of 13 different elements (Al, Ba, Ca, Cr, Cu, La, Mg, Na, Ni, Si, Sr, U and Y) 73% of the spatial variance could be explained based on the wetland coverage and the affinity for organic matter, the latter of which was quantified using thermodynamic modelling. Hence, it was possible to link the large-scale transport patterns of a wide range of metals to fundamental biogeochemical properties. When restraining the analysis to the smaller streams (<10 km2), the explanatory power increased to 88%. For elements such as Na and Si with low affinity for organic matter the decrease in wetland-dominated catchments corresponded closely to the area of mineral soils that had been replaced by peat, indicating that reduced weathering was the main cause of the decrease. For organophilic metals the decrease in wetland-dominated catchments was even greater, suggesting that there also was an accumulation of these metals in the peat. This was confirmed by investigating the distribution of radionuclides in local mire, which revealed considerable accumulation of uranium and thorium along the edges of the mire. Based on the inventories of uranium and thorium and their distribution in the peat it was concluded that the mire historically had been a sink for these metals and that it most likely will continue to be so for a long time to come. All and all, wetlands were estimated to decrease the fluxes of metals from the boreal forests to downstream lakes and oceans by 20-40%, depending on how strongly they bind to organic matter.

The aim of this study was to examine the metal contributions to and from the lake Råsvalen, in northern Örebro County in Sweden, from incoming and outgoing streams.  The main questions were: what is the metal concentration in the streams? How large is the metal transport from the streams, do the concentrations imply a risk for the biota, and can any source be identified. Water was sampled at six locations at six occasions with two-week intervals. The water samples were analyzed for total metal concentrations. The results were compared to environmental quality standards; EQS and from Swedish environmental protection agency suggested class limits. The concentrations were also compared to deviations from background levels and estimations of risk for biological effects. Metal transport was calculated from the average measured concentrations and water flow data from SMHI. Data from sediment analysis in 1989 was included for information about temporal trends. Zn, Cu, and Pb had concentrations above the limits and therefore they could have an effect on biota in water. Zn had a maximum concentration of 39 µg/l in Storån, which is 12 times higher than limits. Pb was found in a maximum concentration of 4,3 µg/l in Hammarskogsån downstream, which is more than twice as high as EQS. Cu was found in concentration of 6,1 µg/l in Storån downstream. Comparison with estimations of biological effects showed no or little effects except for Pb in Hammarskogsån downstream and Storån showing moderate risk. The stream that contributes most to metal influx is Storån. The stream stands for >80% of the total estimated metal transport. Possible sources could be historical mining, and higher levels upstream may contribute to the metal levels found in Storån.

Metal pollution has been a great challenge in most industrialized countries as a result of waste generated from industrial activities being introduced into the environment. Unicellular green algae have been considered a potential biological tool for bioremediation of metal pollutants due to its metal sequestration properties. However, methods for further improving unicellular green algae metal sequestration by manipulating metal uptake and tolerance in unicellular green algae have not been studied in detail. In this study, a family metal transport protein named MTP1 - MTP4 from C. reinhardtii were screened by yeast heterologous expression for metal transport activity. MTP1 was able to strongly rescue the Zn and Co sensitivity of the zrc1cot1 strain, MTP3 could weakly mediate Zn and Co growth, but MTP2 and MTP4 appeared to have no Zn or Co tolerance activity. MTP2, MTP3 and MTP4 but not MTP1 could strongly rescue the Mn sensitivity of the pmr1 strain. When MTP4 was over-expressed in C. reinhardtii the strain showed a significant increase in Cd tolerance compared to the wild type, but no significant difference in Mn tolerance and uptake. AtHMA4 a Zn2+ and Cd2+ transporter from the plant Arabidopsis thaliana, which is a member of the Heavy Metal ATPase family, was also expressed in C. reinhardtii. HMA4 full length and C-terminal tail expression strains were screened for Zn and Cd tolerance and uptake. Both sets of strains showed a significant increase in Cd and Zn tolerance and uptake compared to the wild type. Metal tolerance and uptake was compared between the genetically engineered C. reinhardtii strains and unicellular green algal strains that are naturally adapted to metal tolerance which were P. hussi, P. kessleri, and C. luteoviridis. Results showed significant increase in Zn and Cd tolerance and uptake in the natural strains compared to the engineered strains. Therefore in addition to genetically engineered strains, naturally adapted strains could also be used as tools for effective metal bioremediation and pollutant treatment.

We investigated the transport of dissolved arsenic in groundwater, surface water and the hyporheic zone in a stream-aquifer system influenced by an abandoned arsenopyrite mine. Mine tailing piles consisting of a host of arsenic-bearing minerals including arsenopyrite and scorodite remain adjacent to the stream and represent a continuous source of arsenic. Arsenic loads from the stream, springs, and groundwater were quantified at the study reach on nine dates from January to August 2005 and a mass-balance approach was used to determine hyporheic retention. Arsenic loading from the groundwater was the dominate source of arsenic to the stream, while loads from springs represented a substantial proportion of the total arsenic load during spring. Arsenic loads in surface and groundwater were significantly elevated during summer. Elevated temperatures during summer may lead to increased arsenic loading by increasing dissolution rate of arsenic source minerals and/or increases in microbially-mediated dissolution processes. The hyporheic zone was shown to be retaining arsenic in the upstream-most sub-reach. Retention most likely occurs through the sorption of dissolved arsenic onto hyporheic sediments. In downstream sub-reaches, hyporheic sediments are derived from mine-tailing piles which have high arsenic content. The hyporheic zone in these sub-reaches was shown to be releasing dissolved arsenic. The historic influence of mining activity has resulted in multiple sources of arsenic to the stream which has increased arsenic contamination of the surface waters. / Master of Science

No description available.

Heavy metal pollutants are a worldwide concern. It causes negative effects on aquatic organisms and human health. Heavy metals concentration and transport of copper, zinc and cadmium were investigated in high and low flow conditions in Trönningeån River, southern Sweden. A total of 33 surface water samples collected from the river and Kistingebäcken tributary were analyzed. Concentration (high to low) of heavy metals in the Trönningeån   river and its tributary were- copper(Cu) > zinc (Zn) > cadmium (Cd). The concentration of Copper was found to be high in low flow condition whereas in the case of zinc, high concentrations were found in both the flows (high and low). Study further showed that, the tributary has high pH and conductivity. And finally, the study concluded that there is high concentration and transport of heavy metals in the above-mentioned industrial tributary.

Le but du projet était d'acquérir des connaissances plus approfondies sur les mécanismes impliqués dans l'absorption et l'accumulation des éléments traces métalliques (ETM) par les plantes et dans la résistance à leurs effets toxiques. Arabidopsis halleri, peut vivre sur des sols fortement pollués par des ETM et a la capacité d'accumuler des concentrations extraordinairement élevées de ces métaux dans les tissus aériens. A. halleri est utilisée comme espèce modèle pour étudier les mécanismes biologiques et moléculaires impliqués dans la tolérance et l'accumulation de fortes concentrations de métaux dans le sol. Cette espèce est caractérisée par une tolérance élevée à de fortes concentrations de Cd et Zn. Elle est également capable d'accumuler de très grandes quantités de Zn dans les pousses, appelées hyperaccumulateurs de Zn, et également de Cd dans certaines populations, appelées hyperaccumulateurs de Cd. Arabidopsis arenosa, une espèce étroitement liée à A. halleri, montre également une tolérance élevée aux deux métaux mais est signalée comme non-hyperaccumulatrice. Dans le sud de la Pologne, les deux espèces peuvent être observées sur les mêmes sites pollués. Les expériences ont été conçues pour étudier la variabilité inter et intra spécifique entre les populations métallisées (M) et non métallisées (NM) d'A. halleri et A. arenosa afin d'identifier les paramètres physiologiques et les gènes responsables du comportement contrasté de leur adaptation à la contamination métallique. L'accumulation des ETM, l'activité de l'appareil photosynthétique et le contenu pigmentaire ont été analysés dans les populations M et NM d'A. halleri et A. arenosa, poussant dans leur habitat naturel. Des expériences hydroponiques ont également été réalisées afin de déterminer les différences physiologiques entre les populations étudiées dans des conditions contrôlées. De plus, le niveau d'expression des gènes impliqués dans l'absorption des métaux, le transport radial, la translocation et la détoxification a également été étudié. Mes résultats montrent que la population M de A. arenosa est aussi tolérante aux métaux lourds que la population M de A. halleri. Il a été démontré que la population M de A. arenosa hyperaccumule le Cd et le Zn. De plus, les deux espèces diffèrent dans la réponse de l'appareil photosynthétique lorsqu'elles sont exposées à des concentrations élevées de Cd et de Zn, ce qui suggère des mécanismes différents impliqués dans l'homéostasie des métaux. Le niveau d'expression des gènes a montré des différences non seulement entre les espèces mais aussi entre les populations. Le type de traitement (Cd ou Zn) a également eu un effet différent sur toutes les populations en ce qui concerne certains gènes. Les résultats acquis dans ce projet comblent les lacunes dans les caractéristiques physiologiques des espèces d'A. halleri à la fois sur le terrain et dans des conditions hydroponiques contrôlées. La caractérisation physiologique approfondie de la population M de A. arenosa a identifié cette population comme un nouvel hyperaccumulateur de Cd et Zn. Ces résultats peuvent servir de base à de futures études sur le phénomène d'hyperaccumulation. De plus, l'observation de l'expression contrastée des gènes impliqués dans l'absorption, la translocation et la détoxification des métaux entre A. halleri et A. arenosa suggère que A. arenosa pourrait être un bon nouveau modèle pour étudier l'homéostasie et la tolérance aux métaux chez les plantes. / Doctorat en Sciences agronomiques et ingénierie biologique / info:eu-repo/semantics/nonPublished

Strand et al. (2002) discovered small metal alloy grains rich in Cu, Co, and Sn (maximum size 150 µm) in vesicles of lava from Kilauea Volcano. These alloys are also found in basaltic rocks of several Italian volcanoes. To better understand the origin of these metal-rich grains, bombs from Stromboli Volcano were examined. Two bomb types were collected from Stromboli: pumiceous bombs and scoriaceous bombs. Bulk rock trace element geochemistry indicates that there are no significant differences in Cu, Co, or Sn (the three major components of the metal alloys) between the pumiceous and scoriaceous bombs. Comparison of olivine melt inclusion and matrix glass concentrations from these rocks shows that the pumiceous bombs are more primitive (melt inclusions: MgO 2.7-5.8 wt. %; matrix glass: MgO 5.1-6.50 wt. %) and are more S-rich (melt inclusions: maximum 0.13 wt. %; matrix glass: maximum 0.06 wt. % ) than the scoriaceous bombs. The melt inclusions and matrix glass in the scoriaceous bombs are more evolved (melt inclusions: MgO 3.0-4.3 wt. %; matrix glass: MgO 2.7-3.7 wt. %) and are S-poor (melt inclusions: maximum 0.06 wt. %; matrix glass: b.d.l. ). However, Cl concentrations in melt inclusions and matrix glass are more similar for both bomb types. Metal alloys were counted in thin section for each sample. The crystallized interiors of the bombs contain more metal grains than the glassy exteriors. Pumiceous bombs (from more primitive, S-rich magma) contain more metal grains of a larger size than the scoriaceous bombs (from more fractionated, S-poor magma). This indicates that S (and Cl) are probable transport ligands for the metals in the alloys. As S (and Cl) move through the glass of an erupted cooling bomb, they complex with volatile chalcophile metals (Cu, Co, and Sn). These vapor-phase metal sulfides and chlorides move to inflating vesicles. Here the sulfide and chloride complexes become reduced and metal alloys condense, as S and Cl escape as gas. Non-degassed primitive magma may provide more S (but not necessarily more metals) to create the higher abundance of alloys hosted by the vesicles of the pumiceous bombs.