Transfer of Sulfur from IscS to IscU during Fe/S Cluster Assembly

The cysteine desulfurase enzymes NifS and IscS provide sulfur for the biosynthesis of Fe/S proteins. NifU and IscU have been proposed to serve as template or scaffold proteins in the initial Fe/S cluster assembly events, but the mechanism of sulfur transfer from NifS or IscS to NifU or IscU has not been elucidated. We have employed [35S]cysteine radiotracer studies to monitor sulfur transfer between IscS and IscU from Escherichia coli and have used direct binding measurements to investigate interactions between the proteins. IscS catalyzed transfer of 35S from [35S]cysteine to IscU in the absence of additional thiol reagents, suggesting that transfer can occur directly and without involvement of an intermediate carrier. Surface plasmon resonance studies and isothermal titration calorimetry measurements further revealed that IscU binds to IscS with high affinity (Kd ~2 µM) in support of a direct transfer mechanism. Transfer was inhibited by treatment of IscU with iodoacetamide, and 35S was released by reducing reagents, suggesting that transfer of persulfide sulfur occurs to cysteinyl groups of IscU. A deletion mutant of IscS lacking C-terminal residues 376-413 (IscSDelta 376-413) displayed cysteine desulfurase activity similar to the full-length protein but exhibited lower binding affinity for IscU, decreased ability to transfer 35S to IscU, and reduced activity in assays of Fe/S cluster assembly on IscU. The findings with IscSDelta 376-413 provide additional support for a mechanism of sulfur transfer involving a direct interaction between IscS and IscU and suggest that the C-terminal region of IscS may be important for binding IscU

Structural analysis of the GH43 enzyme Xsa43E from Butyrivibrio proteoclasticus

The rumen of dairy cattle can be thought of as a large, stable fermentation vat and as such it houses a large and diverse community of microorganisms. The bacterium Butyrivibrio proteoclasticus is a representative of a significant component of this microbial community. It is a xylan-degrading organism whose genome encodes a large number of open reading frames annotated as fibre-degrading enzymes. This suite of enzymes is essential for the organism to utilize the plant material within the rumen as a fuel source, facilitating its survival in this competitive environment. Xsa43E, a GH43 enzyme from B. proteoclasticus, has been structurally and functionally characterized. Here, the structure of selenomethionine-derived Xsa43E determined to 1.3 Å resolution using single-wavelength anomalous diffraction is reported. Xsa43E possesses the characteristic five-bladed β-propeller domain seen in all GH43 enzymes. GH43 enzymes can have a range of functions, and the functional characterization of Xsa43E shows it to be an arabinofuranosidase capable of cleaving arabinose side chains from short segments of xylan. Full functional and structural characterization of xylan-degrading enzymes will aid in creating an enzyme cocktail that can be used to completely degrade plant material into simple sugars. These molecules have a range of applications as starting materials for many industrial processes, including renewable alternatives to fossil fuels

Hsc66 substrate specificity is directed toward a discrete region of the iron-sulfur cluster template protein IscU

Hsc66 and Hsc20 comprise a specialized chaperone system important for the assembly of iron-sulfur clusters in Escherchia coli. Only a single substrate, the Fe/S template protein IscU, has been identified for the Hsc66/Hsc20 system, but the mechanism by which Hsc66 selectively binds IscU is unknown. We have investigated Hsc66 substrate specificity using phage display and a peptide array of IscU. Screening of a heptameric peptide phage display library revealed that Hsc66 prefers peptides with a centrally located Pro-Pro motif. Using a cellulose-bound peptide array of IscU we determined that Hsc66 interacts specifically with a region (residues 99-103, LPPVK) that is invariant among all IscU family members. A synthetic peptide (ELPPVKIHC) corresponding to IscU residues 98-106 behaves in a similar manner to native IscU, stimulating the ATPase activity of Hsc66 with similar affinity as IscU, preventing Hsc66 suppression of bovine rhodanese aggregation, and interacting with the peptide-binding domain of Hsc66. Unlike native IscU, however, the synthetic peptide is not bound by Hsc20 and does not synergistically stimulate Hsc66 ATPase activity with Hsc20. Our results indicate that Hsc66 and Hsc20 recognize distinct regions of IscU and further suggest that Hsc66 will not bind LPPVK motifs with high affinity in vivo unless they are in the context of native IscU and can be directed to Hsc66 by Hsc20

Educational materials review

The goal of this project will be: 1. Establish an Educational Materials Review Committee at the District level of the Department of Health and Environmental Control, representative of the disciplines and programs in the District by March 30, 1999 -- A. Develop the process by which materials are submitted to the DMRC for approval -- B. Develop appropriate review criteria by which the materials will be evaluated. -- C. Design the process by which materials, once approved, will be made available to staff

A sub-basin scale dust plume source frequency inventory for southern Africa, 2005-2008

We present a dust plume source inventory for southern Africa. In order to locate and track the local, short-lived plume events, source and frequency data have been derived from Meteosat Second Generation (MSG) thermal infrared composite data (4 km data using 8.7, 10.8, and 12.0 µm) and Moderate Resolution Imaging Spectroradiometer (MODIS) visible composite data (0.25 km data utilizing 0.620 – 0.670 µm, 0.545 – 0.565 µm, and 0.459 – 0.479 µm). Between January 2005 and December 2008, a total of 328 distinct daytime dust plumes more than 10 km in length were detected. These plumes were attributed to 101 distinct point sources, consisting largely of ephemeral inland lakes, coastal pans as well as dry river valleys in Namibia, Botswana, and South Africa. These data also provided sub-basin scale source observations for large basins such as Etosha and Makgadikgadi Pans

Book Reviews

Review of the following books: Revolutionary Outlaws: Ethan Allen and the Struggle for Independence on the Early American Frontier by Michael A. Bellesiles; The Salem Witch Crisis by Larry Gragg; Hard Times, Hard Men: Maine and the Irish, 1830-1860 by James H. Mundy; Keepers of Our Past: Local Historical Writing in the United States, 1820s-1930s by David J. Russo; Woodsmen and Whigs: Historic Images of Bangor, Maine by Abigail Ewing Zelz and Marily Zoidis; Bethel, Maine: An Illustrated History by Randall H. Bennett; Dearest Father: The Civil War Letters of Lt. Frank Dickerson, a Son of Belfast, Maine by H. Draper Hunt

Successfully translating conservation research into practice and policy: concluding thoughts

In the Anthropocene, when our environment is changing rapidly and the windows of opportunity for action to prevent further biodiversity loss are narrow, conservation researchers are increasingly encouraged to think and operate beyond the traditional approaches of producing peer-reviewed papers and presenting results to other members of the research community. Indeed, the perception that researchers belong in their ivory tower, from which they deliver evidence for others to interpret, disseminate and use in decision-making, is thankfully now widely recognised as outdated. The rise of fake news, a deliberate lack of consideration for scientific evidence, and changes to the ways of assessing the value of researchers’ work probably all play a role in supporting this shift in perception. Moreover, for many researchers, the prospect of their work ‘making a difference’ and having an impact on wider society is at least as great a motivation for doing research as generating new knowledge, however interesting that may be

Making a difference in conservation: linking science and policy

amie Gundry’s dramatic image of a white-tailed eagle (Haliaeetus albicilla) on the cover of this book reflects the twisting changes in fortune experienced by this species, with a revival that can be attributed to a successful interplay of science, policy and practice. White-tailed eagles were historically much more widely distributed than they are today (Yalden, 2007), once breeding across much of Europe, but by the early twentieth century the species was extinct across much of western and southern Europe. The main cause of its decline was persecution by farmers and shepherds, who considered the eagles a threat to their livestock, but, along with other raptors, white-tailed eagles were also seriously affected by DDT in the 1960s and 1970s, which had disastrous effects on the breeding success of remaining populations

Lack of Evidence for the Direct Activation of Endothelial Cells by Adult Female and Microfilarial Excretory-Secretory Products

Lymphangiectasia (dilation of the lymphatic vessel (LV)) is pathognomonic for lymphatic filariasis. In both infected humans and animal models of infection, lymphangiectasia is not restricted to the site of the worm nest, but is found along the infected vessel. These observations argue that soluble products secreted by the worm could be mediating this effect by activating the lymphatic endothelial cells (LEC) lining the vessel. We tested the ability of filarial Excretory-Secretory products to activate LECs, but were unable to detect a direct effect of the Excretory-Secretory products on the activation of LEC as assessed by a variety of approaches including cellular proliferation, cell surface molecule expression and cytokine and growth factor production (although other mediators used as positive controls did induce these effects). Collectively, these results do not support the hypothesis that Excretory-Secretory products directly activate LECs

Defining the key wintering habitats in the Sahel for declining African-Eurasian migrants using expert assessment

SummaryThe Sahel in West Africa is a major wintering area for many western Palearctic migrants. The breeding populations of many of these have declined over the past 50 years. However, there have been few intensive field studies on migrant ecology in the Sahel and these were generally within a very restricted area. Consequently our knowledge of the distribution of species within this extensive area and the habitat associations of these species is limited. Understanding these habitat associations is essential for the effective conservation management of populations. We brought together a group of experts and consulted a wider group by email to assess the main Sahelian habitat types used by 68 African-Eurasian migrant bird species. Those species that showed strongest declines during 1970–1990 were associated with more open habitats than those newly declining during 1990–2000, when declining species were associated with habitats with more shrubs and trees. Populations of species that winter in the Sahel are generally stable or increasing now as rainfall has increased and is now near the long-term average for the Sahel. Those which use the Sahel only as a staging area are, in many cases, in rapid decline at present.We would like to thank Andy Clements, Paul Donald, Lincoln Fishpool and Mike Mortimore for contributing to the workshop and Peter Jones, Ian Newton, Volker Salewski, Tim Wacher, Eddy Wymenga and Leo Zwarts for useful comments by email on draft habitat importance scores. This study was funded by the Newton Trust and the Cambridge Conservation Initiative Collaborative Fund, supported by Arcadia. WJS is funded by Arcadia.This is the accepted manuscript of a paper published in Bird Conservation International, Volume 24, Issue 04, December 2014, pp 477-491, DOI: http://dx.doi.org/10.1017/S0959270913000531, Published online: 24 February 201