Molecular organization of the postsynaptic specialization

A specific set of molecules including glutamate receptors is targeted to the postsynaptic specialization of excitatory synapses in the brain, gathering in a structure known as the postsynaptic density (PSD). Synaptic targeting of glutamate receptors depends on interactions between the C-terminal tails of receptor subunits and specific PDZ domain-containing scaffold proteins in the PSD. These scaffold proteins assemble a specialized protein complex around each class of glutamate receptor that functions in signal transduction, cytoskeletal anchoring, and trafficking of the receptors. Among the glutamate receptor subtypes, the N-methyl-d-aspartate receptor is relatively stably integrated in the PSD, whereas the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor moves in and out of the postsynaptic membrane in highly dynamic fashion. The distinctive cell biological behaviors of N-methyl-d-aspartate and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors can be explained by their differential interactions with cytoplasmic proteins.

The multidomain protein Trio binds the LAR transmembrane tyrosine phosphatase, contains a protein kinase domain, and has separate rac-specific and rho-specific guanine nucleotide exchange factor domains.

rho-like GTP binding proteins play an essential role in regulating cell growth and actin polymerization. These molecular switches are positively regulated by guanine nucleotide exchange factors (GEFs) that promote the exchange of GDP for GTP. Using the interaction-trap assay to identify candidate proteins that bind the cytoplasmic region of the LAR transmembrane protein tyrosine phosphatase (PT-Pase), we isolated a cDNA encoding a 2861-amino acid protein termed Trio that contains three enzyme domains: two functional GEF domains and a protein serine/threonine kinase (PSK) domain. One of the Trio GEF domains (Trio GEF-D1) has rac-specific GEF activity, while the other Trio GEF domain (Trio GEF-D2) has rho-specific activity. The C-terminal PSK domain is adjacent to an Ig-like domain and is most similar to calcium/calmodulin-dependent kinases, such as smooth muscle myosin light chain kinase which similarly contains associated Ig-like domains. Near the N terminus, Trio has four spectrin-like repeats that may play a role in intracellular targeting. Northern blot analysis indicates that Trio has a broad tissue distribution. Trio appears to be phosphorylated only on serine residues, suggesting that Trio is not a LAR substrate, but rather that it forms a complex with LAR. As the LAR PTPase localizes to the ends of focal adhesions, we propose that LAR and the Trio GEF/PSK may orchestrate cell-matrix and cytoskeletal rearrangements necessary for cell migration.

Vienna, Austria, 1868 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Plan von Wien : nach den neuesten Quellen gezeichnet und in Kupfer gestochen, von Anton Mück; schrift [gef] I. Bonhammer; lith inst. F. Köke in Wien. It was published by L. T. Neumann in [1868]. Scale [ca. 1:11,500]. Map in German. Covers Vienna, Austria. The image inside the map neatline is georeferenced to the surface of the earth and fit to the MGI 3-Degree Gauss Kruger coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, railroads and stations, drainage, built-up areas and selected buildings, fortification, ground cover, parks, city districts, and more. Includes index and inset: Umgebung von Wien. This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

Neues bürgerliches Kochbuch oder gründliche Anweisung zur Kochkunst für alle Stände

hrsg. von Christian Heinrich Steinbeck, Mundkoch bei Ihro Hoheit der verwittweten Frau Fürstin Reuß-Gera

Vorschriften für den Unterricht in den Waffenübungen der Königlich-bayerischen Artillerie

Lithogr.

Die Pfarrkirche der Augsb. Conf. Verwandten [Augsburger Konfession] zu Hermannstadt

beschrieben von Samuel Möckesch, Cand. d. Theologie

Cloning and expression of the large zebrafish protocadherin gene, Fat

The cadherin superfamily members play an important role in mediating cell-cell contact and adhesion (Takeichi, M., 1991. Cadherin cell adhesion receptors as a morphogenetic regulator. Science 251, 1451-1455). A distinct subfamily, neither belonging to the classical or protocadherins includes Fat, the largest member of the cadherin super-family. Fat was originally identified in Drosophila. Subsequently, orthologues of Fat have been described in man (Dunne, J., Hanby, A. M., Poulsom, R., Jones, T. A., Sheer, D., Chin, W. G., Da, S. M., Zhao, Q., Beverley, P. C., Owen, M. J., 1995. Molecular cloning and tissue expression of FAT, the human homologue of the Drosophila fat gene that is located on chromosome 4q34-q35 and encodes a putative adhesion molecule. Genomics 30, 207-223), rat (Ponassi, M., Jacques, T. S., Ciani, L., ffrench, C. C., 1999. Expression of the rat homologue of the Drosophila fat tumour suppressor gene. Mech. Dev. 80, 207-212) and mouse (Cox, B., Hadjantonakis, A. K., Collins, J. E., Magee, A. I., 2000. Cloning and expression throughout mouse development of mfat 1, a homologue of the Drosophila tumour suppressor gene fat [In Process Citation]. Dev. Dyn. 217, 233-240). In Drosophila, Fat has been shown to play an important role in both planar cell polarity and cell boundary formation during development. In this study we describe the characterization of zebrafish Fat, the first non-mammalian, vertebrate Fat homologue to be identified. The Fat protein has 64% amino acid identity and 80% similarity to human FAT and an identical domain structure to other vertebrate Fat proteins. During embryogenesis fat mRNA is expressed in the developing brain, specialised epithelial surfaces the notochord, ears, eyes and digestive tract, a pattern similar but distinct to that found in mammals. (c) 2005 Elsevier B.V. All rights reserved.

The Digital Observatory for Protected Areas (DOPA) as a support tool for protected area planning, financing and reporting

The Joint Research Centre (JRC) of the European Commission has developed, in consultation with many partners, the DOPA as a global reference information system to support decision making on protected areas (PAs) and biodiversity conservation. The DOPA brings together the World Database on Protected Areas with other reference datasets on species, habitats, ecoregions, threats and pressures, to deliver critical indicators at country level and PA level that can inform gap analyses, PA planning and reporting. These indicators are especially relevant to Aichi Targets 11 and 12, and have recently contributed to CBD country dossiers and capacity building on these targets. DOPA also includes eConservation, a new module that provides a means to share and search information on conservation projects, and thus allows users to see “who is doing what where”. So far over 5000 projects from the World Bank, GEF, CEPF, EU LIFE Programme, CBD LifeWeb Initiative and others have been included, and these projects can be searched in an interactive mapping interface based on criteria such as location, objectives, timeframe, budget, the organizations involved, target species etc. This seminar will provide an introduction to DOPA and eConservation, highlight how these services are used by the CBD and others, and include ample time for discussion.

The Expression of Rap6 in the Adult Mouse Brain and Early Mouse Embryonic Development

The previously identified RAP6 (Rab5 activating protein 6) was associated with plasma membrane mediated endocytosis and contains a Rab5 guanine nucleotide exchange factor (GEF) domain. RAP6 has been shown to act a Ras activating protein (GAP) domain. The identification of RAP6 and its crucial role in both receptors mediated endocytosis and fluid phase endocytosis presents the opportunity to investigate its role in murine embryonic development and in the adult brain. To confirm and characterize the presence of RAP6 during embryonic development and in the adult brain, the current study examined the expression of both the RGD and the Vps9 domains of RAP6 through in situ hybridization. We present an extensive evaluation of the expression for both RAP6 domains through in situ hybridization of 12.5 and 14.5 weeks old C67 mouse embryos and adult C67 mouse brain. The current study confirms the presence of both RAP6 domains and presents an extensive evaluation its expression in embryonic development and the adult brain. These data together support the role of RAP6 in receptor mediated endocytosis and fluid phase endocytosis relevant active during murine embryonic development and adult brain processes.

Intracellular Signaling and Trafficking in Cancer: Role of Rab5-GTPase in Migration and Invasion of Breast Cells

Metastasis is characterized pathologically by uncontrolled cell invasion, proliferation, migration and angiogenesis. Steroid hormones, such as estrogen, and growth factors, which include insulin growth factor I/II (IGF-1/IGF-2) therapy has been associated with most if not all of the features of metastasis. It has been determined that IGF-1 increases cell survival of cancer cells and potentiate the effect of E2 and other ligand growth factors on breast cancer cells. However not much information is available that comprehensively expounds on the roles of insulin growth factor receptor (IGFR) and Rab GTPases may play in breast cancer. The latter, Rab GTPases, are small signaling molecules and critical in the regulation of many cellular processes including cell migration, growth via the endocytic pathway. This research involves the role of Rab GTPases, specifically Rab5 and its guanine exchange factors (GEFs), in the promotion of cancer cell migration and invasion. Two important questions abound: Are IGFR stimulation and downstream effect involved the endocytic pathway in carcinogenesis? What role does Rab5 play in cell migration and invasion of cancer cells? The hypothesis is that growth factor signaling is dependent on Rab5 activity in mediating the aggressiveness of cancer cells. The goal is to demonstrate that IGF-1 signaling is dependent on Rab5 function in breast cancer progression. Here, the results thus far, have shown that while activation of Rab5 may mediate increased cell proliferation, migration and invasion in breast cancer cells, the Rab5 GEF, RIN1 interacts with the IGFR thereby facilitating migration and invasion activities in breast cells. Furthermore, endocytosis of the IGFR in breast cancer cells seems to be caveolin dependent as the data has shown. This taken together, the data shows that IGF-1 signaling in breast cancer cells relies on IGF-1R phosphorylation, caveolae internalization and sequestration to the early endosome RIN1 function and Rab5 activation.

Function of the PDLIM2 protein in oncogenic Wnt signalling pathways

Aberrant regulation of the Wnt signalling pathway is a recurrent theme in cancer biology. Hyper activation due to oncogenic mutations and paracrine activity has been found in both colon cancer and breast cancer, and continues to evolve as a central mechanism in oncogenesis. PDLIM2, a cytoskeletal PDZ protein, is an IGF-1 regulated gene that is highly expressed in cancer cell lines derived from metastatic tumours. Suppression of PDLIM2 inhibits polarized cell migration, reverses the Epithelial to Mesenchymal transition (EMT) phenotype, suppresses the transcription of β-catenin target genes, and regulates gene expression of key transcription factors in EMT. This thesis investigates the mechanism by which PDLIM2 contributes to the maintenance of Wnt signalling in cancer cells. Here we show that PDLIM2 is a critical regulator of the Wnt pathway by regulating β-catenin at the adherens juctions, as also its transcriptional activity by the interaction of PDLIM2 with TCF4 at the nucleus. Evaluation of PDLIM2 in macrophages and co-culture studies with cancer cells and fibroblasts showed the influence exerted on PDLIM2 by paracrine cues. Thus, PDLIM2 integrates cytoskeleton signalling with gene expression by modulating the Wnt signalling pathway and reconciling microenvironmental cues with signals in epithelial cells. Negative correlation of mRNA and protein levels in the triple negative breast cancer cell BT549 suggests that PDLIM2 is part of a more complex mechanism that involves transcription and posttranslational modifications. GST pulldown studies and subsequent mass spectrometry analysis showed that PDLIM2 interacts with 300 proteins, with a high biological function in protein biosynthesis and Ubiquitin/proteasome pathways, including 13 E3 ligases. Overall, these data suggest that PDLIM2 has two distinct functions depending of its location. Located at the cytoplasm mediates cytoskeletal re-arrangements, whereas at the nucleus PDLIM2 acts as a signal transduction adaptor protein mediating transcription and ubiquitination of key transcription factors in cancer development.

An investigation of the functional role of GRAB in the context of Rab3, Rab8 and Rab11

Rab GTPases are the largest family of the Ras superfamily and are key regulators of membrane trafficking within the cell. There are over 60 members of the Rab family which localise to specific membrane compartments and interact with effector proteins to regulate membrane trafficking processes, such as vesicle formation, vesicle trafficking within the cell and fusion with an acceptor compartment. Multiple effector proteins have been identified for many Rabs, some of which can interact with more than one Rab to link their function at a specific membrane location or to link them together in a Rab activation cascade. Rabin8 is one such protein which is an effector for Rab11a and a Guanine nucleotide Exchange Factor (GEF) for Rab8a. Rabin8 participates in a conserved Rab activation cascade which is critical in the formation of primary cilia. Data presented in this thesis has shown that GRAB interacts with Rab3a, Rab8a, Rab11a and Rab11b in a nucleotide dependent manner. Furthermore, the minimal interacting regionbetween these proteins has been investigated. The functional outcome of GRAB knockdown has also been examined and data in this thesis highlights the phenotypic outcome.

Characterization of ALDH1A1 as a novel tumorigenic target mediating TAZ-induced lung tumorigenesis and stem cell phenotypes

Recent studies suggest that lung cancer stem cells (CSCs) may play major roles in lung cancer development, metastasis and drug resistance. Therefore, identification of lung CSC drivers may provide promising targets for lung cancer. TAZ (transcriptional co-activator with PDZ-binding motif) is a transcriptional co-activator and key downstream effector of the Hippo pathway, which plays critical roles in various biological processes. TAZ has been shown to be overexpressed in non-small cell lung cancer (NSCLC) and involved in tumorigenicity of lung epithelial cells. However, whether TAZ is a driver for lung CSCs and tumor formation in vivo is unknown. In addition, the molecular mechanism underlying TAZ-induced lung tumorigenesis remains to be determined. In this study, we provided evidence that constitutively active TAZ (TAZ-S89A) is a driver for lung tumorigenesis in vivo in mice and formation of lung CSC. Oncogenes upregulated in TAZ-overexpressing cells were identified with further validation. The most dramatically activated gene, Aldh1a1 (Aldehyde dehydrogenase 1 family member a1), a well-established CSC marker, showed that TAZ induces Aldh1a1 transcription by activating its promoter activity through interaction with the transcription factor TEA domain (TEAD) family member. Most significantly, inhibition of ALDH1A1 with its inhibitor A37 or CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) gene knockout in lung cancer cells suppressed lung tumorigenic and CSC phenotypes in vitro, and tumor formation in mice in vivo. In conclusion, this study identified TAZ as a novel inducer of lung CSCs and the first transcriptional activator of the stem cell marker ALDH1A1. Most significantly, we identified ALDH1A1 as a critical meditator of TAZ-induced tumorigenic and CSC phenotypes in lung cancer. Our studies provided preclinical data for targeting of TAZ-TEAD-ALDH1A1 signaling to inhibit CSC-induced lung tumorigenesis and drug resistance in the future.

Those Who Belong and Those Who Don’t: Physical and Mental Borders in Europe

Europe promotes migration and mobility but new or ‘different’ Europeans are still stigmatised and marginalised in our societies. Today, neither refugee status nor citizenship can tear down the mental borders between people who inhabit the same cities or neighbourhoods. Sociology professor Nira Yuval-Davis writes about the meaning of borders, and how they make us differentiate between ‘us’ and ‘the other’ in our daily lives.