The crystal structure of the Sgt1-Skp1 complex: the link between Hsp90 and both SCF E3 ubiquitin ligases and kinetochores

The essential cochaperone Sgt1 recruits Hsp90 chaperone activity to a range of cellular factors including SCF E3 ubiquitin ligases and the kinetochore in eukaryotes. In these pathways Sgt1 interacts with Skp1, a small protein that heterodimerizes with proteins containing the F-box motif. We have determined the crystal structure of the interacting domains of Saccharomyces cerevisiae Sgt1 and Skp1 at 2.8 Å resolution and validated the interface in the context of the full-length proteins in solution. The BTB/POZ domain of Skp1 associates with Sgt1 via the concave surface of its TPR domain using residues that are conserved in humans. Dimerization of yeast Sgt1 occurs via an insertion that is absent from monomeric human Sgt1. We identify point mutations that disrupt dimerization and Skp1 binding in vitro and find that the interaction with Skp1 is an essential function of Sgt1 in yeast. Our data provide a structural rationale for understanding the phenotypes of temperature-sensitive Sgt1 mutants and for linking Skp1-associated proteins to Hsp90-dependent pathways

Soil structure and greenhouse gas production differences between row and interrow positions under no-tillage

No-tillage in Brazil is an effi cient agricultural system that improves crop productivity whilst controlling erosion caused to the soil by degradation. However, there is some concern regarding soil compaction. Our objective was to determine whether the function of soil structure in sustaining crop growth was dependent on row and interrow positions in long-term no-tillage. We took soil samples from a fi eld in a commercial farm under long-term no-tillage since 1979 on a clayey Oxisol in Southern Brazil. We assessed soil physical quality using the revised Peerlkamp technique and measured bulk density, air-fi lled porosity and air permeability of intact soil cores. Samples were incubated to assess in vitro N2O and CO2 production. The soil physical and structural properties showed consistent differences between interrow and row positions, where the properties measured were more favorable. The revised Peerlkamp technique proved as effi cient as quantitative parameters in discriminating treatment differences. Overall, soil physical conditions in the interrow were less favourable than in the row. Pore continuity did not vary as regards position. This may explain why row position did not infl uence in vitro N2O and CO2 production. Soil physical quality under no-tillage system is enhanced, at least in the short term, by superficial disturbances in the row as a result of the action of the coulters of the no-tillage seeder

The merits of the Visual Evaluation of Soil Structure method (VESS) for assessing soil physical quality in the remote, undeveloped regions of the Amazon basin

The Visual Evaluation of Soil Structure (VESS) is a straightforward and logistically simple method for characterising and scoring soil structural and physical quality, ideally suited to evaluate and monitor soil degradation in remote and undeveloped areas. The research presented here tested for the first time the feasibility of using VESS in the Amazon basin, under the specialised land uses and soils (Yellow Oxisol and "Terra Preta de Índio") of the region, and its relation with quantitative soil indicators. The evaluated areas, which had never been subjected to mechanisation, fertilisation nor tillage, were "Terra Preta de Índio"/ Anthropogenic Dark Earth; Regenerating Forest; Slash and Burn; Pasture; and Pristine Forest. The results showed that the quantitative indicators were less sensitive at revealing signs of degradation than VESS and that VESS brought to light evidence of historic land use change and limitations to crop productivity. VESS was significantly correlated with soil resistance to penetration. However, VESS had difficulty capturing possible low water-holding capacity and surface sealing, but the hands on approach to VESS allowed the user to identify these problems, despite not being listed in the reference chart. Overall, VESS was a more integrated soil quality indicator, exposing more aspects of soil functionality than the quantitative indicators, it was also logistically easier to perform making it ideal for tracking soil degradation and structural quality in similarly challenging situations. However, more research is required to fully enable VESS to capture structural quality in 'sandified' soils, caused by the slash and burn method widely used in the Amazon region

Cyclin A is a prognostic indicator in early stage breast cancer with and without tamoxifen treatment

Overexpression of G1-S regulators cyclin D1 or cyclin A is frequently observed in breast cancer and is also to result in ligand-independent activation of oestrogen receptor in vitro. This might therefore, provide a mechanism for failure of tamoxifen treatment. We examined by immunohistochemical staining the effect of deregulation of these, and other cell cycle regulators on tamoxifen treatment in a group of 394 patients with early stage breast cancer. In univariate analysis, expression of cyclin A, Neu, Ki-67 index, and lack of OR expression were significantly associated with worse prognosis. When adjusted by the clinical model (for lymph node status, age, performance status, T-classification, grade, prior surgery, oestrogen receptor status and tamoxifen use), only overexpression of cyclin A and Neu were significantly associated with worse prognosis with hazard ratios of, respectively, 1.709 (P=0.0195) and 1.884 (P=0.0151). Overexpression of cyclin A was found in 86 out of the 201 OR-positive cases treated with tamoxifen, and was the only independent marker associated with worse prognosis (hazard ratio 2.024, P=0.0462). In conclusion, cyclin A is an independent predictor of recurrence of early stage breast cancer and is as such a marker for response in patients treated with tamoxifen

Transcriptome response of high- and low-light-adapted Prochlorococcus strains to changing iron availability

Prochlorococcus contributes significantly to ocean primary productivity. The link between primary productivity and iron in specific ocean regions is well established and iron-limitation of Prochlorococcus cell division rates in these regions has been demonstrated. However, the extent of ecotypic variation in iron metabolism among Prochlorococcus and the molecular basis for differences is not understood. Here, we examine the growth and transcriptional response of Prochlorococcus strains, MED4 and MIT9313, to changing iron concentrations. During steady-state, MIT9313 sustains growth at an order-of-magnitude lower iron concentration than MED4. To explore this difference, we measured the whole-genome transcriptional response of each strain to abrupt iron starvation and rescue. Only four of the 1159 orthologs of MED4 and MIT9313 were differentially-expressed in response to iron in both strains. However, in each strain, the expression of over a hundred additional genes changed, many of which are in labile genomic regions, suggesting a role for lateral gene transfer in establishing diversity of iron metabolism among Prochlorococcus. Furthermore, we found that MED4 lacks three genes near the iron-deficiency induced gene (idiA) that are present and induced by iron stress in MIT9313. These genes are interesting targets for studying the adaptation of natural Prochlorococcus assemblages to local iron conditions as they show more diversity than other genomic regions in environmental metagenomic databases.Gordon and Betty Moore FoundationNational Science Foundation (U.S.) (Biological Oceanography)United States. Office of Naval Research (ONR Young Investigator Award)National Science Foundation (U.S.) (Chemical Oceanography)National Science Foundation (U.S.) (Environmental Genomics grants