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https://www.readbyqxmd.com/read/28521592/molecular-perturbation-strategies-to-examine-spatiotemporal-features-of-rho-gef-and-rho-gtpase-activity-in-living-cells
#1
Joachim Goedhart, Jakobus van Unen
Much of our current knowledge of Rho GTPase networks and the regulation by Rho guanine exchange factors (Rho GEFs) and Rho GTPase activating proteins (Rho GAPs) is based on population-based techniques. Over the last decades, technologies that enable single cell analysis with high spatial and temporal resolution have revealed that Rho GTPase activity in cells is regulated on second timescales and at submicrometer length scales. Therefore, perturbation methods with matching spatial and temporal resolution are crucial to further our understanding of Rho GTPase signaling...
May 19, 2017: Small GTPases
https://www.readbyqxmd.com/read/28516903/molecular-mechanism-of-g%C3%AE-i-activation-by-non-gpcr-proteins-with-a-g%C3%AE-binding-and-activating-motif
#2
Alain Ibáñez de Opakua, Kshitij Parag-Sharma, Vincent DiGiacomo, Nekane Merino, Anthony Leyme, Arthur Marivin, Maider Villate, Lien T Nguyen, Miguel Angel de la Cruz-Morcillo, Juan B Blanco-Canosa, Sekar Ramachandran, George S Baillie, Richard A Cerione, Francisco J Blanco, Mikel Garcia-Marcos
Heterotrimeric G proteins are quintessential signalling switches activated by nucleotide exchange on Gα. Although activation is predominantly carried out by G-protein-coupled receptors (GPCRs), non-receptor guanine-nucleotide exchange factors (GEFs) have emerged as critical signalling molecules and therapeutic targets. Here we characterize the molecular mechanism of G-protein activation by a family of non-receptor GEFs containing a Gα-binding and -activating (GBA) motif. We combine NMR spectroscopy, computational modelling and biochemistry to map changes in Gα caused by binding of GBA proteins with residue-level resolution...
May 18, 2017: Nature Communications
https://www.readbyqxmd.com/read/28515233/wntless-sec12-complex-on-er-membrane-regulates-early-wnt-secretory-vesicle-assembly-and-mature-ligand-export
#3
Jiaxin Sun, Shiyan Yu, Xiao Zhang, Catherine Capac, Onyedikachi Aligbe, Timothy Daudelin, Edward M Bonder, Nan Gao
Wntless (Wls) transports Wnt molecules for secretion, however the cellular mechanism underlying the initial assembly of Wnt secretory vesicles is still not fully defined. Using proteomic and mutagenic analyses of the mammalian Wls, we report a mechanism for formation of early Wnt secretory vesicle on ER membrane. Wls forms a complex with SEC12, an ER membrane-localized GEF activator of SAR1 small GTPase. Compared to palmitoylation-deficient Wnt molecules, binding of mature Wnt to Wls increases Wls-SEC12 interaction and promotes Wls association with SAR1, the key activator of the COPII machinery...
May 17, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28515142/flow-induced-endothelial-cell-alignment-requires-the-rhogef-trio-as-a-scaffold-protein-to-polarize-active-rac1-distribution
#4
Jeffrey Kroon, Niels Heemskerk, Martin J T Kalsbeek, Vivian de Waard, Jos van Rijssel, Jaap D van Buul
Endothelial cells line the lumen of the vessel wall and are exposed to flow. In linear parts of the vessel, the endothelial cells experience laminar flow, resulting in endothelial cell alignment in the direction of flow, thereby protecting the vessel wall from inflammation and permeability. In order for endothelial cells to align, they undergo rapid remodeling of the actin cytoskeleton by local activation of the small GTPase Rac1. However, if sustained and local activation of Rac1 is required for long-term flow-induced cell alignment is not clear...
May 17, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28512143/diverse-roles-of-guanine-nucleotide-exchange-factors-in-regulating-collective-cell-migration
#5
Assaf Zaritsky, Yun-Yu Tseng, M Angeles Rabadán, Shefali Krishna, Michael Overholtzer, Gaudenz Danuser, Alan Hall
Efficient collective migration depends on a balance between contractility and cytoskeletal rearrangements, adhesion, and mechanical cell-cell communication, all controlled by GTPases of the RHO family. By comprehensive screening of guanine nucleotide exchange factors (GEFs) in human bronchial epithelial cell monolayers, we identified GEFs that are required for collective migration at large, such as SOS1 and β-PIX, and RHOA GEFs that are implicated in intercellular communication. Down-regulation of the latter GEFs differentially enhanced front-to-back propagation of guidance cues through the monolayer and was mirrored by down-regulation of RHOA expression and myosin II activity...
May 16, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28507547/deletion-of-dock10-in-b-cells-results-in-normal-development-but-a-mild-deficiency-upon-in-vivo-and-in-vitro-stimulations
#6
Natalija Gerasimčik, Minghui He, Marisa A P Baptista, Eva Severinson, Lisa S Westerberg
We sought to identify genes necessary to induce cytoskeletal change in B cells. Using gene expression microarray, we compared B cells stimulated with interleukin-4 (IL-4) and anti-CD40 antibodies that induce B cell spreading, cell motility, tight aggregates, and extensive microvilli with B cells stimulated with lipopolysaccharide that lack these cytoskeletal changes. We identified 84 genes with 10-fold or greater expression in anti-CD40 + IL-4 stimulated B cells, one of these encoded the guanine nucleotide exchange factor (GEF) dedicator of cytokinesis 10 (Dock10)...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28505490/heat-induced-temperature-dysregulation-and-seizures-in-dravet-syndrome-gefs-gabrg2-q390x-mice
#7
Timothy A Warner, Zhong Liu, Robert L Macdonald, Jing-Qiong Kang
It has been established that febrile seizures and its extended syndromes like generalized epilepsy with febrile seizures (FS) plus (GEFS+) and Dravet syndrome have been associated with mutations especially in SCN1A and GABRG2 genes. In patients, the onset of FS is likely due to the combined effect of temperature and inflammation in genetically vulnerable individuals because fever is often associated with infection. Much effort has been spent to understand the mechanisms underlying fever induction of seizures...
April 30, 2017: Epilepsy Research
https://www.readbyqxmd.com/read/28489960/swap70-is-a-universal-gef-like-adapter-for-tethering-actin-to-phagosomes
#8
Maksim V Baranov, Natalia H Revelo, Daniëlle R J Verboogen, Martin Ter Beest, Geert van den Bogaart
We recently identified a key role for SWAP70 as the tethering factor stabilizing F-actin filaments on the surface of phagosomes in human dendritic cells by interacting both with Rho-family GTPases and the lipid phosphatidylinositol (3,4)-bisphosphate. In this study, we aimed to investigate whether this role of SWAP70 was general among immune phagocytes. Our data reveal that SWAP70 is recruited to early phagosomes of macrophages and dendritic cells from both human and mouse. The putative inhibitor of SWAP70 sanguinarine blocked phagocytosis and F-actin polymerization, supporting a key role for SWAP70 in phagocytosis as demonstrated previously with knock-down...
May 10, 2017: Small GTPases
https://www.readbyqxmd.com/read/28487380/egfr-signals-through-a-dock180-mlk3-axis-to-drive-glioblastoma-cell-invasion
#9
Sean Misek, Jian Chen, Laura Schroeder, Chotirat Rattanasinchai, Ashley Sample, Jann N Sarkaria, Kathleen A Gallo
A hallmark of glioblastoma (GBM) tumors is their highly invasive behavior. Tumor dissemination into surrounding brain tissue is responsible for incomplete surgical resection, and subsequent tumor recurrence. Identification of targets that control GBM cell dissemination is critical for developing effective therapies to treat GBM. A majority of GBM tumors have dysregulated EGFR signaling, due most frequently to EGFR amplification or the presence of a constitutively active EGFRvIII mutant. Mixed lineage kinase 3 (MLK3) is a mitogen-activated protein kinase kinase kinase (MAP3K) that can activate multiple MAPK pathways...
May 9, 2017: Molecular Cancer Research: MCR
https://www.readbyqxmd.com/read/28487361/biochemical-characterization-of-purified-mammalian-arl13b-indicate-that-it-is-an-atypical-gtpase-and-arl3-guanine-nucleotide-exchange-factor-gef
#10
Anna A Ivanova, Tamara Caspary, Nicholas T Seyfried, Duc M Duong, Andrew B West, Zhiyong Liu, Richard A Kahn
Primary cilia play central roles in signaling during metazoan development. Several key regulators of ciliogenesis and ciliary signaling are mutated in humans, resulting in a number of ciliopathies, including Joubert Syndrome (JS). ARL13B is a ciliary GTPase with at least three missense mutations identified in JS patients. ARL13B is a member of the ARF family of regulatory GTPases, but is atypical in having a non-homologous, C-terminal domain of ~20 kDa and at least one key residue difference in the consensus GTP binding motifs...
May 9, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28475102/the-small-gtpases-ras-and-rheb-studied-by-multidimensional-nmr-spectroscopy-structure-and-function
#11
REVIEW
Miriam Schöpel, Veena Nambiar Potheraveedu, Thuraya Al-Harthy, Raid Abdel-Jalil, Rolf Heumann, Raphael Stoll
Ras GTPases are key players in cellular signalling because they act as binary switches. These states manifest through toggling between an active (GTP-loaded) and an inactive (GDP-loaded) form. The hydrolysis and replenishing of GTP is controlled by two additional protein classes: GAP (GTPase-activating)- and GEF (Guanine nucleotide exchange factors)-proteins. The complex interplay of the proteins is known as the GTPase-cycle. Several point mutations of the Ras protein deregulate this cycle. Mutations in Ras are associated with up to one-third of human cancers...
May 1, 2017: Biological Chemistry
https://www.readbyqxmd.com/read/28475097/the-small-gtpases-ras-and-rheb-studied-by-multidimensional-nmr-spectroscopy-structure-and-function
#12
Miriam Schöpel, Veena Nambiar Potheraveedu, Thuraya Al-Harthy, Raid Abdel-Jalil, Rolf Heumann, Raphael Stoll
Ras GTPases are key players in cellular signalling because they act as binary switches. These states manifest through toggling between an active (GTP-loaded) and an inactive (GDPloaded) form. The hydrolysis and replenishing of GTP is controlled by two additional protein classes: GAP (GTPase-activating)- and GEF (Guanine nucleotide exchange factors)-proteins. The complex interplay of the proteins is known as the GTPase-cycle. Several point mutations of the Ras protein deregulate this cycle. Mutations in Ras are associated with up to one third of human cancers...
February 11, 2017: Biological Chemistry
https://www.readbyqxmd.com/read/28472098/t1r3-homomeric-sweet-taste-receptor-regulates-adipogenesis-through-g%C3%AE-s-mediated-microtubules-disassembly-and-rho-activation-in-3t3-l1-cells
#13
Yosuke Masubuchi, Yuko Nakagawa, Johan Medina, Masahiro Nagasawa, Itaru Kojima, Mark M Rasenick, Takeshi Inagaki, Hiroshi Shibata
We previously reported that 3T3-L1 cells express a functional sweet taste receptor possibly as a T1R3 homomer that is coupled to Gs and negatively regulates adipogenesis by a Gαs-mediated but cAMP-independent mechanism. Here, we show that stimulation of this receptor with sucralose or saccharin induced disassembly of the microtubules in 3T3-L1 preadipocytes, which was attenuated by overexpression of the dominant-negative mutant of Gαs (Gαs-G226A). In contrast, overexpression of the constitutively active mutant of Gαs (Gαs-Q227L) as well as treatment with cholera toxin or isoproterenol but not with forskolin caused disassembly of the microtubules...
2017: PloS One
https://www.readbyqxmd.com/read/28452363/mechanism-of-sos-pr-domain-autoinhibition-revealed-by-single-molecule-assays-on-native-protein-from-lysate
#14
Young Kwang Lee, Shalini T Low-Nam, Jean K Chung, Scott D Hansen, Hiu Yue Monatrice Lam, Steven Alvarez, Jay T Groves
The guanine nucleotide exchange factor (GEF) Son of Sevenless (SOS) plays a critical role in signal transduction by activating Ras. Here we introduce a single-molecule assay in which individual SOS molecules are captured from raw cell lysate using Ras-functionalized supported membrane microarrays. This enables characterization of the full-length SOS protein, which has not previously been studied in reconstitution due to difficulties in purification. Our measurements on the full-length protein reveal a distinct role of the C-terminal proline-rich (PR) domain to obstruct the engagement of allosteric Ras independently of the well-known N-terminal domain autoinhibition...
April 28, 2017: Nature Communications
https://www.readbyqxmd.com/read/28452129/studies-on-the-bioactivities-of-ace-inhibitory-peptides-with-phenylalanine-c-terminus-using-3d-qsar-molecular-docking-and-in-vitro-evaluation
#15
Chunyan Qi, Guimei Lin, Rong Zhang, Wenjuan Wu
3D-QSAR, molecular docking and activity evaluation were used to study the bioactivities of ACE-inhibitory peptides with phenylalanine C-terminus. Both CoMFA (Q(2) =0.773, R(2) =0.992) and CoMSIA (Q(2) =0.664, R(2) =0.990) models were constructed. According to the established models, four novel potent ACE-inhibitory tripeptides GEF, VEF, VRF, and VKF were synthesized. The IC50 values were respectively determined to be 13 μM, 23 μM, 5 μM, and 11 μM by in vitro evaluation. The results show good agreement with the predicted values...
April 27, 2017: Molecular Informatics
https://www.readbyqxmd.com/read/28448737/involvement-of-arhgef10-gef-for-rhoa-in-rab6-rab8-mediating-membrane-traffic
#16
Satoshi Shibata, Yui Teshima, Kenta Niimi, Shinobu Inagaki
Small GTPases play crucial roles in the maintenance of a homeostatic environment and appropriate movements of the cell. In these processes, the direct or indirect interaction between distinct small GTPases could be required for regulating mutual signaling pathways. In our recent study, ARHGEF10, known as a guanine nucleotide exchange factor (GEF) for RhoA, was indicated to interact with Rab6A and Rab8A, which are known to function in the exocytotic pathway, and colocalized with these Rabs at exocytotic vesicles...
April 27, 2017: Small GTPases
https://www.readbyqxmd.com/read/28444222/feronia-receptor-kinase-at-the-crossroad-of-hormone-signaling-and-stress-responses
#17
Hongdong Liao, Renjie Tang, Xin Zhang, Sheng Luan, Feng Yu
Plant receptor-like kinases (RLKs) are involved in nearly all aspects of plant life including growth, development and stress response. Recent studies show that FERONIA (FER), a CrRLK1L subfamily member, is a versatile regulator of cell expansion and serves as a signaling node mediating crosstalks among multiple phytohormones. As a receptor for the RALF (Rapid Alkalinization Factor) peptide ligand, FER triggers a downstream signaling cascade that leads to a rapid cytoplasmic calcium increase and inhibition of cell elongation in plants...
April 19, 2017: Plant & Cell Physiology
https://www.readbyqxmd.com/read/28442598/ethylene-regulates-differential-growth-via-big-arf-gef-dependent-post-golgi-secretory-trafficking-in-arabidopsis
#18
Kristoffer Jonsson, Yohann Boutté, Rajesh Kumar Singh, Delphine Gendre, Rishikesh Bhalerao
During early seedling development, the shoot apical meristem is protected from damage as the seedling emerges from soil by the formation of apical hook. Hook formation requires differential growth across the epidermis below the meristem in the hypocotyl. The plant hormones ethylene and auxin play key roles during apical hook development by controlling differential growth. We provide genetic and cell biological evidence for the role of ADP-ribosylation factor 1 (ARF1)-GTPase and its effector ARF-guanine-exchange factors (GEF)s of the BFA-inhibited GEF (BIG) family and GNOM in ethylene- and auxin-mediated control of hook development...
April 24, 2017: Plant Cell
https://www.readbyqxmd.com/read/28442529/coinheritance-of-novel-mutations-in-scn1a-causing-gefs-and-in-kdm6a-causing-kabuki-syndrome-in-a-family
#19
Jisun Kim, Cha Gon Lee
Because the differentiation between phenotypic expansion and blended phenotypes is not clear, the mixed phenotypes of blended rare genetic diseases make diagnosis difficult. We describe a family with the co-existence and co-segregation of generalized epilepsy with febrile seizures plus (GEFS+) and Kabuki syndrome (KS). The proband, a 7-year-old male, presented with GEFS+, dysmorphic facial features, short stature, developmental delay, and intellectual disability. Two novel missense mutations: p.G325A in the KDM6A gene responsible for KS and p...
March 2017: Annals of Clinical and Laboratory Science
https://www.readbyqxmd.com/read/28439033/genome-wide-identification-of-resd-nsrr-and-fur-binding-in-bacillus-subtilis-during-anaerobic-fermentative-growth-by-in-vivo-footprinting
#20
Onuma Chumsakul, Divya P Anantsri, Tai Quirke, Taku Oshima, Kensuke Nakamura, Shu Ishikawa, Michiko M Nakano
Upon oxygen limitation, the Bacillus subtilis ResE sensor kinase and its cognate ResD response regulator play primary roles in transcriptional activation of genes functioning in anaerobic respiration. The nitric oxide (NO)-sensitive NsrR repressor controls transcription to support nitrate respiration. In addition, the ferric uptake repressor (Fur) can modulate transcription under anaerobic conditions. However, whether these controls are direct or indirect has been investigated only in a gene-specific manner...
April 24, 2017: Journal of Bacteriology
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