The cysteine-rich frizzled domain of Frzb-1 is required and sufficient for modulation of Wnt signaling

Convincing evidence has accumulated to identify the Frizzled proteins as receptors for the Wnt growth factors. In parallel, a number of secreted frizzled-like proteins with a conserved N-terminal frizzled motif have been identified. One of these proteins, Frzb-1, binds Wnt-1 and Xwnt-8 proteins and antagonizes Xwnt-8 signaling in Xenopus embryos. Here we report that Frzb-1 blocks Wnt-1 induced cytosolic accumulation of β-catenin, a key component of the Wnt signaling pathway, in human embryonic kidney cells. Structure/function analysis reveals that complete removal of the frizzled domain of Frzb-1 abolishes the Wnt-1/Frzb-1 protein interaction and the inhibition of Wnt-1 mediated axis duplication in Xenopus embryos. In contrast, removal of the C-terminal portion of the molecule preserves both Frzb-Wnt binding and functional inhibition of Wnt signaling. Partial deletions of the Frzb-1 cysteine-rich domain maintain Wnt-1 interaction, but functional inhibition is lost. Taken together, these findings support the conclusion that the frizzled domain is necessary and sufficient for both activities. Interestingly, Frzb-1 does not block Wnt-5A signaling in a Xenopus functional assay, even though Wnt-5A coimmunoprecipitates with Frzb-1, suggesting that coimmunoprecipitation does not necessarily imply inhibition of Wnt function.

The Interaction of Activated Integrin Lymphocyte Function-associated Antigen 1 with Ligand Intercellular Adhesion Molecule 1 Induces Activation and Redistribution of Focal Adhesion Kinase and Proline-rich Tyrosine Kinase 2 in T Lymphocytes

Integrin receptors play a central role in the biology of lymphocytes, mediating crucial functional aspects of these cells, including adhesion, activation, polarization, migration, and signaling. Here we report that induction of activation of the β2-integrin lymphocyte function-associated antigen 1 (LFA-1) in T lymphocytes with divalent cations, phorbol esters, or stimulatory antibodies is followed by a dramatic polarization, resulting in a characteristic elongated morphology of the cells and the arrest of migrating lymphoblasts. This cellular polarization was prevented by treatment of cells with the specific tyrosine kinase inhibitor genistein. Furthermore, the interaction of the activated integrin LFA-1 with its ligand intercellular adhesion molecule 1 induced the activation of the cytoplasmic tyrosine kinases focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (PYK-2). FAK activation reached a maximum after 45 min of stimulation; in contrast, PYK-2 activation peaked at 30 min, declining after 60 min. Upon polarization of lymphoblasts, FAK and PYK-2 redistributed from a diffuse localization in the cytoplasm to a region close to the microtubule-organizing center in these cells. FAK and PYK-2 activation was blocked when lymphoblasts were pretreated with actin and tubulin cytoskeleton-interfering agents, indicating its cytoskeletal dependence. Our results demonstrate that interaction of the β2-integrin LFA-1 with its ligand intercellular adhesion molecule 1 induces remodeling of T lymphocyte morphology and activation and redistribution of the cytoplasmic tyrosine kinases FAK and PYK-2.

The solution structure of the Raf-1 cysteine-rich domain: a novel ras and phospholipid binding site.

The Raf-1 protein kinase is the best-characterized downstream effector of activated Ras. Interaction with Ras leads to Raf-1 activation and results in transduction of cell growth and differentiation signals. The details of Raf-1 activation are unclear, but our characterization of a second Ras-binding site in the cysteine-rich domain (CRD) and the involvement of both Ras-binding sites in effective Raf-1-mediated transformation provides insight into the molecular aspects and consequences of Ras-Raf interactions. The Raf-1 CRD is a member of an emerging family of domains, many of which are found within signal transducing proteins. Several contain binding sites for diacylglycerol (or phorbol esters) and phosphatidylserine and are believed to play a role in membrane translocation and enzyme activation. The CRD from Raf-1 does not bind diacylglycerol but interacts with Ras and phosphatidylserine. To investigate the ligand-binding specificities associated with CRDs, we have determined the solution structure of the Raf-1 CRD using heteronuclear multidimensional NMR. We show that there are differences between this structure and the structures of two related domains from protein kinase C (PKC). The differences are confined to regions of the CRDs involved in binding phorbol ester in the PKC domains. Since phosphatidylserine is a common ligand, we expect its binding site to be located in regions where the structures of the Raf-1 and PKC domains are similar. The structure of the Raf-1 CRD represents an example of this family of domains that does not bind diacylglycerol and provides a framework for investigating its interactions with other molecules.

The Las Campanas/Anglo-Australian Telescope Rich Cluster Survey - III. Spectroscopic studies of X-ray bright galaxy clusters at z similar to 0.1

We present the analysis of the spectroscopic and photometric catalogues of 11 X-ray luminous clusters at 0.07 < z < 0.16 from the Las Campanas/Anglo-Australian Telescope Rich Cluster Survey. Our spectroscopic data set consists of over 1600 galaxy cluster members, of which two-thirds are outside r(200). These spectra allow us to assign cluster membership using a detailed mass model and expand on our previous work on the cluster colour-magnitude relation ( CMR) where membership was inferred statistically. We confirm that the modal colours of galaxies on the CMR become progressively bluer with increasing radius d( B - R)/dr(p) = - 0.011 +/- 0.003 and with decreasing local galaxy density d( B - R)/dlog ( Sigma)= - 0.062 +/- 0.009. Interpreted as an age effect, we hypothesize that these trends in galaxy colour should be reflected in mean H delta equivalent width. We confirm that passive galaxies in the cluster increase in Hd line strength as dH delta/dr(p) = 0.35 +/- 0.06. Therefore, those galaxies in the cluster outskirts may have younger luminosity-weighted stellar populations; up to 3 Gyr younger than those in the cluster centre assuming d( B - R)/dt = 0.03 mag per Gyr. A variation of star formation rate, as measured by [ O II]lambda 3727 angstrom, with increasing local density of the environment is discernible and is shown to be in broad agreement with previous studies from the 2dF Galaxy Redshift Survey and the Sloan Digital Sky Survey. We divide our spectra into a variety of types based upon the MORPHs classification scheme. We find that clusters at z similar to 0.1 are less active than their higher-redshift analogues: about 60 per cent of the cluster galaxy population is non-star forming, with a further 20 per cent in the post-starburst class and 20 per cent in the currently active class, demonstrating that evolution is visible within the past 2 - 3 Gyr. We also investigate unusual populations of blue and very red non-star forming galaxies and we suggest that the former are likely to be the progenitors of galaxies which will lie on the CMR, while the colours of the latter possibly reflect dust reddening. We show that the cluster galaxies at large radii consist of both backsplash ones and those that are infalling to the cluster for the first time. We make a comparison to the field population at z similar to 0.1 and examine the broad differences between the two populations. Individually, the clusters show significant variation in their galaxy populations which we suggest reflects their recent infall histories.

(Table 1, page 126) Concentration of elements in Fe and Mn rich pisoliths (nodules) from Lake Storsjoen, South Norway

A detailed description of the ores of Lake Storsjoen was given by Vogt J. H. L., 1915 who pointed out that the ores may be divided into two principal types; first, iron ore with 2% or less of manganese (ex: Ertemalm), and, second, ores with manganese contents of up to 30% (ex: Korinter). The iron-rich ore sometimes occurs as a conglomerate embedded in manganese-rich ores, clearly demonstrating that two distinctly different precipitates are involved. In the iron-rich ore, a concentric structure is common of which light brown layers of loose, almost dusty material alternate with hard and brittle black layers, the thickness of each being 0.5 mm or less. The analyses presented in this paper seem to demonstrate that the composition of the sedimentary ores of Lake Storsjden could result from fluctuations in the composition of ground waters feeding the lake.