Identification of the Axin and Frat binding region of glycogen synthase kinase-3.

Glycogen synthase kinase-3 (GSK-3) is a key component of several signaling pathways including those regulated by Wnt and insulin ligands. Specificity in GSK-3 signaling is thought to involve interactions with scaffold proteins that localize GSK-3 regulators and substrates. This report shows that GSK-3 forms a low affinity homodimer that is disrupted by binding to Axin and Frat. Based on the crystal structure of GSK-3, we have used surface-scanning mutagenesis to identify residues that differentially affect GSK-3 interactions. Mutations that disrupt Frat and Axin cluster at the dimer interface explaining their effect on homodimer formation. Loss of the Axin binding site blocks the ability of dominant negative GSK-3 to cause axis duplication in Xenopus embryos. The Axin binding site is conserved within all GSK-3 proteins, and its loss affects both cell motility and gene expression in the nonmetazoan, Dictyostelium. Surprisingly, we find no genetic interaction between a non-Axin-binding GSK-3 mutant and T-cell factor activity, arguing that Axin interactions alone cannot explain the regulation of T-cell factor-mediated gene expression.     

The Reactions of Methanimine and Cyanogen with Carbon Monoxide in Prebiotic Molecular Evolution on Earth

A primeval atmosphere is proposedcontaining simple molecules such as formaldehyde, ammonia, carbon monoxide, cyanogen andhydrogen cyanide, which have been detected in space. Chemical reactions aredescribed for the formation ofaziridine-2-one and di-azirine-3-one derivatives aspotential precursors for the original synthesesis of amino-acids, proteins, pyrimidines,purines, nicotinamide and flavin. The reactions have been shown to be kinetically feasiblefrom the overall enthalpy changes in the ZKE approximation at the MP2/6-31G^* level.     

Benthic macrofauna in Tasmanian estuaries: scales of distribution and relationships with environmental variables

Information on the distribution of species richness, faunal density, biomass and estimated productivity of benthic invertebrates in Tasmanian estuaries was quantified at a variety of spatial and temporal scales to assess general hypothesis relating community metrics to such environmental variables as salinity, seagrass biomass and sediment particle size. An associated aim was to assess appropriate scales of investigation for soft-sediment biota distributed in estuaries, including whether patterns identified at individual sites, estuaries, tidal levels or times are likely to have more general relevance. Faunal biomass and productivity varied principally at between-estuary (10 to 1000 km) and replicate-sample (1 m) scales, indicating that these two community metrics were largely responding to estuary-wide effects, such as nutrient loading, and to microhabitat features, rather than to locality characteristics at intermediate scales such as salinity, anoxia or sediment particle size. By contrast, faunal density showed greater response to tidal height (1 to 100 m) and to factors distributed at the locality scale within estuary (10 km) than to factors between estuary. Both faunal density and species richness in estuaries declined over three- and fivefold ranges down the shore from high water mark to the shallow sublittoral, while estimated productivity and biomass showed highest overall levels at low water mark. The greatest component of variance in species richness was associated with tidal height, with variance then distributed approximately evenly between other spatial scales examined. At the low-tide and shallow subtidal levels, species richness, faunal biomass and estimated productivity were all highly correlated with salinity and biomass of macrophytes, whereas faunal density was highly correlated with biomass of macrophytes only. Relationships between environmental and biological variables examined were poorly defined at high tidal levels. Seasonal plus interannual variance was much lower than spatial variance—a clear indication that sampling effort in studies would generally be better directed across a range of localities than for a single locality to be repeatedly investigated over time.     

Butenolide from plant-derived smoke enhances germination and seedling growth of arable weed species

We tested the applicability of the recently identified major germination cue from smoke (a butenolide 3-methyl-2Hfuro[2,3-c]pyran-2-one) on 18 weed species from non-fire prone environments. For the study species we compared the relative effectiveness of alternating temperatures, KNO₃, GA₃, smoke water and the butenolide on germination percentage, germination rate and seedling mass. We found that while smoke stimulated germination in a number of species it also had negative impacts on other species. In addition, the butenolide was effective on the widest range of species in terms of enhancing germination percentage, rate and seedling mass. However, none of the treatments, including butenolide were effective on all species. Our data demonstrate that butenolide may have wide applicability as a germination and seedling growth stimulant irrespective of whether the species come from fire-prone habitats.     

Human killer cells and natural killer cells: distinct subpopulations of Fc receptor-bearing lymphocytes

Unstimulated human peripheral blood lymphocytes were depleted of K cells, which mediate antibody-dependent cellular cytotoxicity (ADCC) without removing NK cells, which mediate natural killing (NK). K cell depletion was achieved by buoyant centrifugation removal of lymphocytes that bound to glutaraldehyde-treated P815-AB cells at high lymphocyte-to-target ratios. Likewise, NK cells were removed with glutaraldehyde-treated K562 cells without removing K cells. Furthermore, both cytotoxic cell populations were observed directly in one agarose single-cell cytotoxic assay (ASCA) using P815-AB and K562 cells simultaneously as target cells. Moreover, the percentage of total cytotoxic cells was equal to the sum of the percentage of K and NK cells observed in separate ASCA. Collectively, these results indicate that K cells and NK cells are distinct subsets of FcR-bearing lymphocytes. One subset, K cells, has more avid Fc receptors (fcR) than NK cells and are 'activated' via thier FcR to kill antibody-coated target cells. The second subset, NK cells, have less avid FcR and are not 'activated' through their FcR to kill antibody-coated target cells.