The Tea4-PP1 landmark promotes local growth by dual Cdc42 GEF recruitment and GAP exclusion.

Cell polarization relies on small GTPases, such as Cdc42, which can break symmetry through self-organizing principles, and landmarks that define the axis of polarity. In fission yeast, microtubules deliver the Tea1-Tea4 complex to mark cell poles for growth, but how this complex activates Cdc42 is unknown. Here, we show that ectopic targeting of Tea4 to cell sides promotes the local activation of Cdc42 and cell growth. This activity requires that Tea4 binds the type I phosphatase (PP1) catalytic subunit Dis2 or Sds21, and ectopic targeting of either catalytic subunit is similarly instructive for growth. The Cdc42 guanine-nucleotide-exchange factor Gef1 and the GTPase-activating protein Rga4 are required for Tea4-PP1-dependent ectopic growth. Gef1 is recruited to ectopic Tea4 and Dis2 locations to promote Cdc42 activation. By contrast, Rga4 is locally excluded by Tea4, and its forced colocalization with Tea4 blocks ectopic growth, indicating that Rga4 must be present, but at sites distinct from Tea4. Thus, a Tea4-PP1 landmark promotes local Cdc42 activation and growth both through Cdc42 GEF recruitment and by creating a local trough in a Cdc42 GAP.

The ubiquitin-like protein LC3 regulates the Rho-GEF activity of AKAP-Lbc.

AKAP-Lbc is a member of the A-kinase anchoring protein (AKAP) family that has been recently associated with the development of pathologies, such as cardiac hypertrophy and cancer. We have previously demonstrated that, at the molecular level, AKAP-Lbc functions as a guanine nucleotide exchange factor (GEF) that promotes the specific activation of RhoA. In the present study, we identified the ubiquitin-like protein LC3 as a novel regulatory protein interacting with AKAP-Lbc. Mutagenesis studies revealed that LC3, through its NH(2)-terminal alpha-helical domain, interacts with two binding sites located within the NH(2)-terminal regulatory region of AKAP-Lbc. Interestingly, LC3 overexpression strongly reduced the ability of AKAP-Lbc to interact with RhoA, profoundly impairing the Rho-GEF activity of the anchoring protein and, as a consequence, its ability to promote cytoskeletal rearrangements associated with the formation of actin stress fibers. Moreover, AKAP-Lbc mutants that fail to interact with LC3 show a higher basal Rho-GEF activity as compared with the wild type protein and become refractory to the inhibitory effect of LC3. This suggests that LC3 binding maintains AKAP-Lbc in an inactive state that displays a reduced ability to promote downstream signaling. Collectively, these findings provide evidence for a previously uncharacterized role of LC3 in the regulation of Rho signaling and in the reorganization of the actin cytoskeleton.

Anchoring of both PKA and 14-3-3 inhibits the Rho-GEF activity of the AKAP-Lbc signaling complex.

A-kinase anchoring proteins (AKAPs) target the cAMP-regulated protein kinase (PKA) to its physiological substrates. We recently identified a novel anchoring protein, called AKAP-Lbc, which functions as a PKA-targeting protein as well as a guanine nucleotide exchange factor (GEF) for RhoA. We demonstrated that AKAP-Lbc Rho-GEF activity is stimulated by the alpha subunit of the heterotrimeric G protein G12. Here, we identified 14-3-3 as a novel regulatory protein interacting with AKAP-Lbc. Elevation of the cellular concentration of cAMP activates the PKA holoenzyme anchored to AKAP-Lbc, which phosphorylates the anchoring protein on the serine 1565. This phosphorylation event induces the recruitment of 14-3-3, which inhibits the Rho-GEF activity of AKAP-Lbc. AKAP-Lbc mutants that fail to interact with PKA or with 14-3-3 show a higher basal Rho-GEF activity as compared to the wild-type protein. This suggests that, under basal conditions, 14-3-3 maintains AKAP-Lbc in an inactive state. Therefore, while it is known that AKAP-Lbc activity can be stimulated by Galpha12, in this study we demonstrated that it is inhibited by the anchoring of both PKA and 14-3-3.

Expression of the Rap1 guanine nucleotide exchange factor, MR-GEF, is altered in individuals with bipolar disorder

In the rodent forebrain GABAergic neurons are generated from progenitor cells that express the transcription factors Dlx1 and Dlx2. The Rap-1 guanine nucleotide exchange factor, MR-GEF, is turned on by many of these developing GABAergic neurons. Expression of both Dlx1/2 and MR-GEF is retained in both adult mouse and human forebrain where, in human, decreased Dlx1 expression has been associated with psychosis. Using in situ hybridization studies we show that MR-GEF expression is significantly down-regulated in the forebrain of Dlx1/2 double mutant mice suggesting that MR-GEF and Dlx1/2 form part of a common signalling pathway during GABAergic neuronal development. We therefore compared MR-GEF expression by in situ hybridization in individuals with major psychiatric disorders (schizophrenia, bipolar disorder, major depression) and control individuals. We observed a significant positive correlation between layers II and IV of the dorso-lateral prefrontal cortex (DLPFC) in the percentage of MR-GEF expressing neurons in individuals with bipolar disorder, but not in individuals with schizophrenia, major depressive disorder or in controls. Since MR-GEF encodes a Rap1 GEF able to activate G-protein signalling, we suggest that changes in MR-GEF expression could potentially influence neurotransmission.

Expression of the guanine nucleotide exchange factor, mr-gef, is regulated during the differentiation of specific subsets of telencephalic neurons

We have performed a screen combining subtractive hybridization with PCR to isolate genes that are regulated when neuroepithelial (NE) cells differentiate into neurons. From this screen, we have isolated a number of known genes that have not previously been associated with neurogenesis, together with several novel genes. Here we report that one of these genes, encoding a guanine nucleotide exchange factor (GEF), is regulated during the differentiation of distinct neuronal populations. We have cloned both rat and mouse GEF genes and shown that they are orthologs of the human gene, MR-GEF, which encodes a GEF that specifically activates the small GTPase, Rap1. We have therefore named the rat gene rat mr-gef (rmr-gef) and the mouse gene mouse mr-gef (mmr-gef). Here, we will collectively refer to these two rodent genes as mr-gef. Expression studies show that mr-gef is expressed by young neurons of the developing rodent CNS but not by progenitor cells in the ventricular zone (VZ). The expression pattern of mr-gef during early telencephalic neurogenesis is strikingly similar to that of GABA and the LIM homeobox gene Lhx6, a transcription factor expressed by GABAergic interneurons generated in the ventral telencephalon, some of which migrate into the cortex during development. These observations suggest that mr-gef encodes a protein that is part of a signaling pathway involved in telencephalic neurogenesis; particularly in the development of GABAergic interneurons.

Der Körper als Gefäß. Eine Studie zur visuellen Anthropologie des Alten Orients

The perception of the human body as container is widespread in cognitive linguistics, psychology and anthropology and is estimated to be universal. But what can we say about the specific context of the Hebrew Bible as well as Ancient-Near-Eastern texts and material culture, and more especially about anthropomorphic vessels in the Levant? Biblical, Egyptian and Mesopotamian texts compare the human body with pottery in order to emphasize its status of having been created (Geschöpflichkeit) on the one hand, and its fragility (Zerbrechlichkeit) on the other. Not in every case does the metaphor refer to an individual. Very often, however, it is used with relation to groups of people (nations) and requires particular caution when it comes to drawing a conclusion about the embodiment. The archaeological-iconographic record demonstrates that the human body, especially the female body, was imagined as a container. The fact that vessels in the shape of female bodies are the majority can partly be explained with the association between container and pregnancy. This essay aims at stimulating the discussion about embodiment in the Ancient Near East, concepts of emotion the body as a container in the Hebrew Bible, and its relation to the material culture.

Abundance of mesozooplankton in the western part of the Black Sea in summer 2005 during Phase 2: Leg1 from the GEF Project

The sampling area was extended to the Western-South area off the Black Sea coast from Kaliakra cape toward the Bosforous. Samples were collected along four transects. The whole dataset is composed of 17 samples (from 10 stations) with data of mesozooplankton species composition abundance and biomass. Sampling for zooplankton was performed from bottom up to the surface at depths depending on water column stratification and the thermocline depth. These data are organized in the "Control of eutrophication, hazardous substances and related measures for rehabilitating the Black Sea ecosystem: Phase 2: Leg I: PIMS 3065". Data Report is not published. Zooplankton samples were collected with vertical closing Juday net,diameter - 36cm, mesh size 150 µm. Tows were performed from surface down to bottom meters depths in discrete layers. Samples were preserved by a 4% formaldehyde sea water buffered solution. Sampling volume was estimated by multiplying the mouth area with the wire length. Mesozooplankton abundance: The collected material was analysed using the method of Domov (1959). Samples were brought to volume of 25-30 ml depending upon zooplankton density and mixed intensively until all organisms were distributed randomly in the sample volume. After that 5 ml of sample was taken and poured in the counting chamber which is a rectangle form for taxomomic identification and count. Large (> 1 mm body length) and not abundant species were calculated in whole sample. Counting and measuring of organisms were made in the Dimov chamber under the stereomicroscope to the lowest taxon possible. Taxonomic identification was done at the Institute of Oceanology by Kremena Stefanova using the relevant taxonomic literature (Mordukhay-Boltovskoy, F.D. (Ed.). 1968, 1969,1972). Taxon-specific abundance: The collected material was analysed using the method of Domov (1959). Samples were brought to volume of 25-30 ml depending upon zooplankton density and mixed intensively until all organisms were distributed randomly in the sample volume. After that 5 ml of sample was taken and poured in the counting chamber which is a rectangle form for taxomomic identification and count. Copepods and Cladoceras were identified and enumerated; the other mesozooplankters were identified and enumerated at higher taxonomic level (commonly named as mesozooplankton groups). Large (> 1 mm body length) and not abundant species were calculated in whole sample. Counting and measuring of organisms were made in the Dimov chamber under the stereomicroscope to the lowest taxon possible. Taxonomic identification was done at the Institute of Oceanology by Kremena Stefanova using the relevant taxonomic literature (Mordukhay-Boltovskoy, F.D. (Ed.). 1968, 1969,1972).

The Rho-GEF Rom2p Localizes to Sites of Polarized Cell Growth and Participates in Cytoskeletal Functions in Saccharomyces cerevisiae

Rom2p is a GDP/GTP exchange factor for Rho1p and Rho2p GTPases; Rho proteins have been implicated in control of actin cytoskeletal rearrangements. ROM2 and RHO2 were identified in a screen for high-copy number suppressors of cik1Δ, a mutant defective in microtubule-based processes in Saccharomyces cerevisiae. A Rom2p::3XHA fusion protein localizes to sites of polarized cell growth, including incipient bud sites, tips of small buds, and tips of mating projections. Disruption of ROM2 results in temperature-sensitive growth defects at 11°C and 37°C. rom2Δ cells exhibit morphological defects. At permissive temperatures, rom2Δ cells often form elongated buds and fail to form normal mating projections after exposure to pheromone; at the restrictive temperature, small budded cells accumulate. High-copy number plasmids containing either ROM2 or RHO2 suppress the temperature-sensitive growth defects of cik1Δ and kar3Δ strains. KAR3 encodes a kinesin-related protein that interacts with Cik1p. Furthermore, rom2Δ strains exhibit increased sensitivity to the microtubule depolymerizing drug benomyl. These results suggest a role for Rom2p in both polarized morphogenesis and functions of the microtubule cytoskeleton.

Daple is a novel non-receptor GEF required for trimeric G protein activation in Wnt signaling

Peer reviewed

Characterization in Drosophila melanogaster of dPDZ-GEF, a Rap GTPase activator

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Characterization in Drosophila melanogaster of dPDZ-GEF, a Rap GTPase activator

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Soil carbon stocks under no-tillage mulch-based cropping systems in the Brazilian Cerrado: An on-farm synchronic assessment

No-tillage mulch-based (NTM) cropping systems have been widely adopted by farmers in the Brazilian savanna region (Cerrado biome). We hypothesized that this new type of management should have a profound impact on soil organic carbon (SOC) at regional scale and consequently on climate change mitigation. The objective of this study was thus to quantify the SOC storage potential of NTM in the oxisols of the Cerrado using a synchronic approach that is based on a chronosequence of fields of different years under NTM. The study consisted of three phases: (1) a farm/cropping system survey to identify the main types of NTM systems to be chosen for the chronosequence; (2) a field survey to identify a homogeneous set of situations for the chronosequence and (3) the characterization of the chronosequence to assess the SOC storage potential. The main NTM system practiced by farmers is an annual succession of soybean (Glycine max)or maize (Zea mays) with another cereal crop. This cropping system covers 54% of the total cultivated area in the region. At the regional level, soil organic C concentrations from NTM fields were closely correlated with clay + silt content of the soil (r(2) = 0.64). No significant correlation was observed (r(2) = 0.07), however, between these two variables when we only considered the fields with a clay + silt content in the 500-700 g kg(-1) range. The final chronosequence of NTM fields was therefore based on a subsample of eight fields, within this textural range. The SOC stocks in the 0-30 cm topsoil layer of these selected fields varied between 4.2 and 6.7 kg C m(-2) and increased on average (r(2) = 0.97) with 0.19 kg C m(-2) year(-1). After 12 years of NTM management, SOC stocks were no longer significantly different from the stocks under natural Cerrado vegetation (p < 0.05), whereas a 23-year-old conventionally tilled and cropped field showed SOC stocks that were about 30% below this level. Confirming our hypotheses, this study clearly illustrated the high potential of NTM systems in increasing SOC storage under tropical conditions, and how a synchronic approach may be used to assess efficiently such modification on farmers` fields, identifying and excluding non desirable sources of heterogeneity (management, soils and climate). (C) 2010 Elsevier B.V. All rights reserved.