Heterotrophic carbon metabolism and energy acquisition in Candidatus Thioglobus singularis strain PS1, a member of the SUP05 clade of marine Gammaproteobacteria

A hallmark of the SUP05 clade of marine Gammaproteobacteria is the ability to use energy obtained from reduced inorganic sulfur to fuel autotrophic fixation of carbon using RuBisCo. However, some SUP05 also have the genetic potential for heterotrophic growth, raising questions about the roles of SUP05 in the marine carbon cycle. We used genomic reconstructions, physiological growth experiments and proteomics to characterize central carbon and energy metabolism in Candidatus Thioglobus singularis strain PS1, a representative from the SUP05 clade that has the genetic potential for autotrophy and heterotrophy. Here, we show that the addition of individual organic compounds and 0.2 μm filtered diatom lysate significantly enhanced the growth of this bacterium. This positive growth response to organic substrates, combined with expression of a complete TCA cycle, heterotrophic pathways for carbon assimilation, and methylotrophic pathways for energy conversion demonstrate strain PS1's capacity for heterotrophic growth. Further, our inability to verify the expression of RuBisCO suggests that carbon fixation was not critical for growth. These results highlight the metabolic diversity of the SUP05 clade that harbours both primary producers and consumers of organic carbon in the oceans and expand our understanding of specific pathways of organic matter oxidation by the heterotrophic SUP05.     

Prochlorococcus extracellular vesicles: molecular composition and adsorption to diverse microbes

Extracellular vesicles are small (~50–200 nm diameter) membrane-bound structures released by cells from all domains of life. While vesicles are abundant in the oceans, their functions, both for cells themselves and the emergent ecosystem, remain a mystery. To better characterize these particles – a prerequisite for determining function – we analysed the lipid, protein, and metabolite content of vesicles produced by the marine cyanobacterium Prochlorococcus. We show that Prochlorococcus exports a diverse array of cellular compounds into the surrounding seawater enclosed within discrete vesicles. Vesicles produced by two different strains contain some materials in common, but also display numerous strain-specific differences, reflecting functional complexity within vesicle populations. The vesicles contain active enzymes, indicating that they can mediate extracellular biogeochemical reactions in the ocean. We further demonstrate that vesicles from Prochlorococcus and other bacteria associate with diverse microbes including the most abundant marine bacterium, Pelagibacter. Together, our data point toward hypotheses concerning the functional roles of vesicles in marine ecosystems including, but not limited to, possibly mediating energy and nutrient transfers, catalysing extracellular biochemical reactions, and mitigating toxicity of reactive oxygen species.     

The DNA-binding domain of human papillomavirus type 18 E1. Crystal structure, dimerization, and DNA binding

High risk types of human papillomavirus, such as type 18 (HPV-18), cause cervical carcinoma, one of the most frequent causes of cancer death in women worldwide. DNA replication is one of the central processes in viral maintenance, and the machinery involved is an excellent target for the design of antiviral therapy. The papillomaviral DNA replication initiation protein E1 has origin recognition and ATP-dependent DNA melting and helicase activities, and it consists of a DNA-binding domain and an ATPase/helicase domain. While monomeric in solution, E1 binds DNA as a dimer. Dimerization occurs via an interaction of hydrophobic residues on a single alpha-helix of each monomer. Here we present the crystal structure of the monomeric HPV-18 E1 DNA-binding domain refined to 1.8-A resolution. The structure reveals that the dimerization helix is significantly different from that of bovine papillomavirus type 1 (BPV-1). However, we demonstrate that the analogous residues required for E1 dimerization in BPV-1 and the low risk HPV-11 are also required for HPV-18 E1. We also present evidence that the HPV-18 E1 DNA-binding domain does not share the same nucleotide and amino acid requirements for specific DNA recognition as BPV-1 and HPV-11 E1.     

A glycoprotein hormone expressed in corticotrophs exhibits unique binding properties on thyroid-stimulating hormone receptor

Corticotroph-derived glycoprotein hormone (CGH), also referred to as thyrostimulin, is a noncovalent heterodimer of glycoprotein hormone alpha 2 (GPHA2) and glycoprotein hormone beta 5 (GPHB5). Here, we demonstrate that both subunits of CGH are expressed in the corticotroph cells of the human anterior pituitary, as well as in skin, retina, and testis. CGH activates the TSH receptor (TSHR); (125)I-CGH binding to cells expressing TSHR is saturable, specific, and of high affinity. In competition studies, unlabeled CGH is a potent competitor for (125)I-TSH binding, whereas unlabeled TSH does not compete for (125)I-CGH binding. Binding and competition analyses are consistent with the presence of two binding sites on the TSHR transfected baby hamster kidney cells, one that can interact with either TSH or CGH, and another that binds CGH alone. Transgenic overexpression of GPHB5 in mice produces elevations in serum T(4) levels, reductions in body weight, and proptosis. However, neither transgenic overexpression of GPHA2 nor deletion of GPHB5 produces an overt phenotype in mice. In vivo administration of CGH to mice produces a dose-dependent hyperthyroid phenotype including elevation of T(4) and hypertrophy of cells within the inner adrenal cortex. However, the distinctive expression patterns and binding characteristics of CGH suggest that it has endogenous biological roles that are discrete from those of TSH.     

Environmental management of a highly impacted,urbanized tropical estuary: rehabilitation and restoration

The principles of the dynamics and interrelationships within the dominant subtropical and tropical Caribbean seagrass community have been studied previously before, during, and after impact. From these and scores of observations of damage and recovery patterns inThalassia ecosystems, a sense of management recovery strategy has emerged. Artificial restoring ofThalassia testudinum seeds into areas cut off from stock (fruit, seeds) appeared feasible on a large scale after the Turkey Point (Biscayne Bay, Miami, Florida) restoration and test sampling throughout North Biscayne Bay. Two large-scale seeding attempts were made; after 11 months they compared favorably with Turkey Point specimens with regard to growth parameters, despite the turbidity and other persistent pollution. Thus, the possible areas in whichThalassia seed restoration can be used has increased to include estuaries of multiple impact still in various stages of recovery after physical and sewage pollution. This technique should be especially useful to “developing” nations where important nearshore fisheries nurseries based onThalassia ecosystems have been heavily damaged and now lie barren. Man's impact on the estuary where seed restoration was attempted includes the following activities: 50% of the bay bottom directly dredged or filled (leaving much unconsolidated sediment); 50 million gallons of domestic waste dumped directly into a low flushing part of the bay for 20 years; seven major causeways transecting the bay, restricting circulation and flushing; two artificial inlets made into navigational channels; freshwater sheet flow drastically changed due to channelization by flood-control canals; urban runoff from a million people entering the bay. Most of the impacts have now abated; however, their long-term effects remain.     

Cultivation of a thermophilic ammonia oxidizing archaeon synthesizing crenarchaeol

The widespread occurrence and diversity of ammonia oxidizing Archaea suggests their contribution to the nitrogen cycle is of global significance. Their distribution appeared limited to low- and moderate-temperature environments until the recent finding of a diagnostic membrane lipid, crenarchaeol, in terrestrial hot springs. We report here the cultivation of a thermophilic nitrifier (' Candidatus Nitrosocaldus yellowstonii'), an autotrophic crenarchaeote growing up to 74°C by aerobic ammonia oxidation. The major core lipid of this archaeon growing at 72°C is crenarchaeol, providing the first direct evidence for its synthesis by a thermophile. These findings greatly extend the upper temperature limit of nitrification and document that the capacity for ammonia oxidation is broadly distributed among the Crenarchaeota .     

Comparison of the role of the foliar sheath in nutrient (ammonium and phosphate) acquisition by the seagrass <Emphasis Type="Italic">Thalassia hemprichii</Emphasis> (Ehrenb.) Aschers. at two different sites on tropical Hainan Island,China

Generally, the foliar sheaths of seagrass contribute a large biomass to the dry weight of plants, and are found to be above-sediment biomass or, sometimes, below-sediment biomass. However, the role of foliar sheaths of seagrass in nutrient uptake has not yet been established. Thus, this study was performed to test whether the growth form of foliar sheaths affects the nutrient uptake properties of the seagrass. Two separate sets of morphotypes of the seagrass Thalassia hemprichii were collected from two different tropical meadows in coastal Hainan Island, China in the South China Sea. Ammonium (NH₄ ⁺) and phosphate (P_i) uptake by solely blades and roots (experiment I), and above and below-sediment tissues (experiment II) of the two sets of specimens were examined in partitioned chambers using laboratory incubations. Curve profiles of the blade and root saturation uptake kinetics were shown to be similar for the two morphotypes of T. hemprichii. However, the above and below-sediment tissues uptake kinetics had different characteristics between the two morphotypes. For plants with above-sediment foliar sheaths, uptake by the above-sediment tissues contributed an important part of the whole plants’ nutrient acquisition. In contrast, for plants with below-sediment foliar sheaths, the contribution of nutrient uptake by above-sediment foliar blade tissues seemed almost negligible. Therefore, the results demonstrated that foliar sheaths of the tropical seagrass T. hemprichii were able to absorb NH₄ ⁺ and P_i. Especially interesting is that the capacity for uptake by robust foliar sheaths growing beneath the sediment was remarkable (we termed this the Zhang–Huang–Thorhaug effect). The role of sheaths in nutrient acquisition found in this study is critical in elucidating seagrass nutrient uptake strategies.     

TRACE METAL CYCLING IN THE U.S. COASTAL ZONE: A SYNTHESIS

This synthesis of trace element research in estuarine communities of the U.S. coastline and the Caribbean provides a summary applicable to the shoreline of the tropical and temperate regions of the world which have mangroves, kelp beds, riverine marshes and seagrass communities. An inventory of sediments and leaf tissues shows Mn and Fe to be the most highly concentrated elements with Hg and Cd present in lowest concentrations. Generally, trace element concentrations in roots are much higher than in leaves and other tissues above the sediment. Tissue to sediment concentration ratios show that Cd is most likely to be bioamplified and that Cu, Hg, Sr and Zn may have relatively high concentration ratios which can exceed unity. A conceptual model was constructed to integrate the forcing functions, compartmental couplings, and dynamics common to these estuarine systems. Seasonality is important for changes in some trace element concentrations in plants and litter. Trace element additions to water or sediment increased certain trace element concentrations in plants and dead organic matter. It is clear that estuarine plant communities serve as living filters of estuarine trace elements. However, increased knowledge of trace element cycling in estuarine systems and relationships between trace element concentrations in plants and the estuarine food chain is needed, particularly food chains to man. There is a need for structured long-term estuarine research to allow direct comparison of results among estuarine study sites, to identify the similarity of population and system processes among estuaries and to define the geographical scale over which estuarine research results may be generalized.