Crystal structure of a monomeric form of SARS-CoV endonuclease nsp15 suggests a role for hexamerization as an allosteric switch

Mature nonstructural protein-15 (nsp15) from the severe acute respiratory syndrome coronavirus (SARS-CoV) contains a novel uridylate-specific Mn2+-dependent endoribonuclease (NendoU). Structure studies of the full-length form of the obligate hexameric enzyme from two CoVs, SARS-CoV and murine hepatitis virus, and its monomeric homologue, XendoU from Xenopus laevis, combined with mutagenesis studies have implicated several residues in enzymatic activity and the N-terminal domain as the major determinant of hexamerization. However, the tight link between hexamerization and enzyme activity in NendoUs has remained an enigma. Here, we report the structure of a trimmed, monomeric form of SARS-CoV nsp15 (residues 28 to 335) determined to a resolution of 2.9 A. The catalytic loop (residues 234 to 249) with its two reactive histidines (His 234 and His 249) is dramatically flipped by approximately 120 degrees into the active site cleft. Furthermore, the catalytic nucleophile Lys 289 points in a diametrically opposite direction, a consequence of an outward displacement of the supporting loop (residues 276 to 295). In the full-length hexameric forms, these two loops are packed against each other and are stabilized by intimate intersubunit interactions. Our results support the hypothesis that absence of an adjacent monomer due to deletion of the hexamerization domain is the most likely cause for disruption of the active site, offering a structural basis for why only the hexameric form of this enzyme is active.

Plant remains as evidence for seasonality of site use in the Mesolithic period

Attempts to identify the seasons during which Mesolithic sites may have been occupied have usually concentrated on evidence from faunal assemblages, while the potential of plant remains has been largely neglected. In this paper the use of plant remains as indicators of seasonality is discussed, especially where they represent the accidental charring of species unlikely to have been deliberately collected and stored. This is illustrated with reference to the early Mesolithic site of Star Carr, North Yorkshire, and discussed in the context of other Mesolithic sites from which assemblages of charred plant remains have been recovered.

New radiocarbon accelerator dates on artefacts from the early Mesolithic site of Star Carr, North Yorkshire

For the well-known early Mesolithic site of Star Carr, dating of organic artefacts by accelerator mass spectrometry (AMS) has been hampered by treatment of bone and antler recovered during the original excavations with preservatives. Some, untreated, artefacts were, however, collected after Clark's excavation in 1950. Four of these artefacts were AMS dated in 1995, but two of the dates were significantly younger than the others, and were questionable due to their low collagen yields. These suspect samples have now been re-analysed, demonstrating that all four artefacts are of similar date. The significance of these dates for the chronology of Star Carr is discussed.

Joined-up archaeology at Old Scatness, Shetland: thin section analysis of the site and hinterland

Excavations on the multi-period settlement at Old Scatness, Shetland have uncovered a number of Iron Age structures with compacted, floor-like layers. Thin section analysis was undertaken in order to investigate and compare the characteristics of these layers. The investigation also draws on earlier analyses of the Iron Age agricultural soil around the settlement and the midden deposits that accumulated within the settlement, to create a 'joined-up' analysis which considers the way material from the settlement was used and then recycled as fertiliser for the fields. Peat was collected from the nearby uplands and was used for fuel and possibly also for flooring. It is suggested that organic-rich floors from the structures were periodically removed and the material was spread onto the fields as fertilisers. More organic-rich material may have been used selectively for fertiliser, while the less organic peat ash was allowed to accumulate in middens. Several of the structures may have functioned as byres, which suggests a prehistoric plaggen system.

Availability of heavy metals for uptake by Salix viminalis on a moderately contaminated dredged sediment disposal site

Extractability of Cd. Cr, Cu, Ni, Pb, and Zn in a dredged sediment disposal site was assessed using single extraction Procedures (H2O; 0.01 M CaCl2; 1 M NH4OAc NH4OAc-EDTA. CaCl2-TEA-DTPA). Only Cd and Zn were Found to exceed statutory threshold values for total content. The field was planted with Salix viminalis "Orm" and accumulation of heavy metals in bark, leaves, roots, and wood was evaluated at seven sampling locations along an observed gradient in texture and pollution. Biomass production was high, ranging from 13.2 to 17.8 t ha(1) y(1) dry weight. Metal accumulation in aboveground Plant parts Was low. amounting to the following annually extracted mass or metals per ha: 5034 g Zn, 83 g Cd. W g Cu. 83 g Pb, 12 g Ni and 6 g Cr. The use of accumulating clones and the use of soil amendments might enhance extraction efficiency in future research. (C) 2005 Elsevier Ltd. All rights reserved.