An allosteric switch controls the procoagulant and anticoagulant activities of thrombin.

Thrombin is an allosteric enzyme existing in two forms, slow and fast, that differ widely in their specificities toward synthetic and natural amide substrates. The two forms are significantly populated in vivo, and the allosteric equilibrium can be affected by the binding of effectors and natural substrates. The fast form is procoagulant because it cleaves fibrinogen with higher specificity; the slow form is anticoagulant because it cleaves protein C with higher specificity. Binding of thrombomodulin inhibits cleavage of fibrinogen by the fast form and promotes cleavage of protein C by the slow form. The allosteric properties of thrombin, which has targeted two distinct conformational states toward its two fundamental and competing roles in hemostasis, are paradigmatic of a molecular strategy that is likely to be exploited by other proteases in the blood coagulation cascade.

Pax5 (BSAP) regulates the murine immunoglobulin 3' alpha enhancer by suppressing binding of NF-alpha P, a protein that controls heavy chain transcription.

The Pax5 transcription factor BSAP (B-cell-specific activator protein) is known to bind to and repress the activity of the immunoglobulin heavy chain 3' alpha enhancer. We have detected an element--designated alpha P--that lies approximately 50 bp downstream of the BSAP binding site 1 and is required for maximal enhancer activity. In vitro binding experiments suggest that the 40-kDa protein that binds to this element (NF-alpha P) is a member of the Ets family present in both B-cell and plasma-cell nuclei. However, in vivo footprint analysis suggests that the alpha P site is occupied only in plasma cells, whereas the BSAP site is occupied in B cells but not in plasma cells. When Pax5 binding to the enhancer in B cells was blocked in vivo by transfection with a triple-helix-forming oligonucleotide an alpha P footprint appeared and endogenous immunoglobulin heavy chain transcripts increased. The triple-helix-forming oligonucleotide also increased enhancer activity of a transfected construct in B cells, but only when the alpha P site was intact. Pax5 thus regulates the 3' alpha enhancer and immunoglobulin gene transcription by blocking activation by NF-alpha P.

Protein export from the nucleus requires the GTPase Ran and GTP hydrolysis.

Nuclei of digitonin-permeabilized cells that had been preloaded with a model transport substrate in a cytosol-dependent import reaction were subsequently incubated to investigate which conditions would result in export of transport substrate. We found that up to 80% of the imported substrate was exported when recombinant human Ran and GTP were present in the export reaction. Ran-mediated export was inhibited by nonhydrolyzable GTP analogs and also by wheat germ agglutinin but was unaffected by a nonhydrolyzable ATP analog. Moreover, a recombinant human Ran mutant that was deficient in its GTPase activity inhibited export. These data indicate that export of proteins from the nucleus requires Ran and GTP hydrolysis but not ATP hydrolysis. We also found that digitonin-permeabilized cells were depleted of their endogenous nuclear Ran, thus allowing detection of Ran as a limiting factor for export. In contrast, most endogenous karyopherin alpha was retained in nuclei of digitonin-permeabilized cells. Unexpectedly, exogenously added, fluorescently labeled Ran, although it accessed the nuclear interior, was found to dock at the nuclear rim in a punctate pattern, suggesting the existence of Ran-binding sites at the nuclear pore complex.

Cell cycle-regulated nuclear import and export of Cdc47, a protein essential for initiation of DNA replication in budding yeast.

The CDC47 gene was isolated by complementation of a cdc47 temperature-sensitive mutant in Saccharomyces cerevisiae and was shown to encode a predicted polypeptide, Cdc47, of 845 aa. Cdc47 belongs to the Cdc46/Mcm family of proteins, previously shown to be essential for initiation of DNA replication. Using indirect immunofluorescence microscopy and subcellular fractionation techniques, we show that Cdc47 undergoes cell cycle-regulated changes in its subcellular localization. At mitosis, Cdc47 enters the nucleus, where it remains until soon after the initiation of DNA replication, when it is rapidly exported back into the cytoplasm. Cdc47 protein levels do not vary with the cell cycle, but expression of CDC47 and nascent synthesis of Cdc47 occur late in the cell cycle, coinciding with mitosis. Together, these results show that Cdc47 is not only imported into the nucleus at the end of mitosis but is also exported back into the cytoplasm at the beginning of S phase. The observation that Cdc47 is exported from the nucleus at the beginning of S phase has important implications for how initiation of DNA replication is controlled.

Climatic and halokinetic controls on alluvial–lacustrine sedimentation during compressional deformation, Andean forearc, northern Chile

Peer reviewed

Illinois Phosphorus Effluent Limits: Minimizing the Impacts Through the Use of Non-point Controls at the Springbrook Water Reclamation Center in Naperville, Illinois

Phosphorus pollution is a major concern in Illinois. Excessive amounts of phosphorus can be detrimental to water bodies. To help control phosphorus, the Illinois Pollution Control Board has proposed phosphorus limits on wastewater treatment facility discharges. If enacted, these limits will have negative impacts on the Springbrook Water Reclamation Center in Naperville, Illinois. To minimize these impacts, Naperville can utilize various non-point controls recommended in this paper to decrease the amount of phosphorus entering into the DuPage River and the Springbrook Water Reclamation Center. While these controls will not reduce levels low enough to totally satisfy limits on phosphorus discharges, they will significantly reduce the treatment costs Naperville will need to expend to meet them and be more environmentally effective.

Terrain Controls and Landscape-Scale Susceptibility Modelling of Active-Layer Detachments, Sabine Peninsula, Melville Island, Nunavut

Modelling the susceptibility of permafrost slopes to disturbance can identify areas at risk to future disturbance and result in safer infrastructure and resource development in the Arctic. In this study, we use terrain attributes derived from a digital elevation model, an inventory of permafrost slope disturbances known as active-layer detachments (ALDs) and generalised additive modelling to produce a map of permafrost slope disturbance susceptibility for an area on northern Melville Island, in the Canadian High Arctic. By examining terrain variables and their relative importance, we identified factors important for initiating slope disturbance. The model was calibrated and validated using 70 and 30 per cent of a data-set of 760 mapped ALDs, including disturbed and randomised undisturbed samples. The generalised additive model calibrated and validated very well, with areas under the receiver operating characteristic curve of 0.89 and 0.81, respectively, demonstrating its effectiveness at predicting disturbed and undisturbed samples. ALDs were most likely to occur below the marine limit on slope angles between 3 and 10° and in areas with low values of potential incoming solar radiation (north-facing slopes).