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https://www.readbyqxmd.com/read/27925692/a-method-to-generate-and-analyze-modified-myristoylated-proteins
#1
Huanyao Gao, Wei Sun, Zhiquan Song, Yanbao Yu, Li Wang, Xian Chen, Qisheng Zhang
Covalent lipid modification of proteins is essential to their cellular localizations and functions. Engineered lipid motif coupled with bio-orthogonal chemistry has been utilized to identify myristoylated or palmitoylated proteins in cells. However, whether modified proteins have similar properties as endogenous ones has not been well investigated mainly due to lack of methods to generate and analyze purified proteins. We have developed a method that utilizes metabolic interference and mass spectrometry to produce and analyze modified, myristoylated small GTPase ADP-ribosylation factor 1 (Arf1)...
December 7, 2016: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/27872253/gtpase-cross-talk-regulates-trappii-activation-of-rab11-homologues-during-vesicle-biogenesis
#2
Laura L Thomas, J Christopher Fromme
Rab guanosine triphosphatases (GTPases) control cellular trafficking pathways by regulating vesicle formation, transport, and tethering. Rab11 and its paralogs regulate multiple secretory and endocytic recycling pathways, yet the guanine nucleotide exchange factor (GEF) that activates Rab11 in most eukaryotic cells is unresolved. The large multisubunit transport protein particle (TRAPP) II complex has been proposed to act as a GEF for Rab11 based on genetic evidence, but conflicting biochemical experiments have created uncertainty regarding Rab11 activation...
November 21, 2016: Journal of Cell Biology
https://www.readbyqxmd.com/read/27848911/phospholipase-d-activity-couples-plasma-membrane-endocytosis-with-retromer-dependent-recycling
#3
Rajan Thakur, Aniruddha Panda, Elise Coessens, Nikita Raj, Shweta Yadav, Sruthi Balakrishnan, Qifeng Zhang, Plamen Georgiev, Bishal Basak, Renu Pasricha, Michael Jo Wakelam, Nicholas T Ktistakis, Padinjat Raghu
During illumination, the light-sensitive plasma membrane (rhabdomere) of Drosophila photoreceptors undergoes turnover with consequent changes in size and composition. However, the mechanism by which illumination is coupled to rhabdomere turnover remains unclear. We find that photoreceptors contain a light-dependent phospholipase D (PLD) activity. During illumination, loss of PLD resulted in an enhanced reduction in rhabdomere size, accumulation of Rab7 positive, rhodopsin1-containing vesicles (RLVs) in the cell body and reduced rhodopsin protein...
November 16, 2016: ELife
https://www.readbyqxmd.com/read/27812735/hygromycin-b-hypersensitive-hhy-mutants-implicate-an-intact-trans-golgi-and-late-endosome-interface-in-efficient-tor1-vacuolar-localization-and-torc1-function
#4
Daniele E Ejzykowicz, Kristopher M Locken, Fiona J Ruiz, Surya P Manandhar, Daniel K Olson, Editte Gharakhanian
Saccharomyces cerevisiae vacuoles are functionally analogous to mammalian lysosomes. Both also serve as physical platforms for Tor Complex 1 (TORC1) signal transduction, the master regulator of cellular growth and proliferation. Hygromycin B is a eukaryotic translation inhibitor. We recently reported on hygromycin B hypersensitive (hhy) mutants that fail to grow at subtranslation inhibitory concentrations of the drug and exhibit vacuolar defects (Banuelos et al. in Curr Genet 56:121-137, 2010). Here, we show that hhy phenotype is not due to increased sensitivity to translation inhibition and establish a super HHY (s-HHY) subgroup of genes comprised of ARF1, CHC1, DRS2, SAC1, VPS1, VPS34, VPS45, VPS52, and VPS54 that function exclusively or inclusively at trans-Golgi and late endosome interface...
November 3, 2016: Current Genetics
https://www.readbyqxmd.com/read/27791132/endocytosis-of-wingless-via-a-dynamin-independent-pathway-is-necessary-for-signaling-in-drosophila-wing-discs
#5
Anupama Hemalatha, Chaitra Prabhakara, Satyajit Mayor
Endocytosis of ligand-receptor complexes regulates signal transduction during development. In particular, clathrin and dynamin-dependent endocytosis has been well studied in the context of patterning of the Drosophila wing disc, wherein apically secreted Wingless (Wg) encounters its receptor, DFrizzled2 (DFz2), resulting in a distinctive dorso-ventral pattern of signaling outputs. Here, we directly track the endocytosis of Wg and DFz2 in the wing disc and demonstrate that Wg is endocytosed from the apical surface devoid of DFz2 via a dynamin-independent CLIC/GEEC pathway, regulated by Arf1, Garz, and class I PI3K...
October 25, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27748959/study-of-ethanol-induced-golgi-disorganization-reveals-the-potential-mechanism-of-alcohol-impaired-n-glycosylation
#6
Carol A Casey, Ganapati Bhat, Melissa S Holzapfel, Armen Petrosyan
BACKGROUND: It is known that ethanol (EtOH) and its metabolites have a negative effect on protein glycosylation. The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. Because previously we have shown that Golgi morphology dictates glycosylation, we examined the effect EtOH administration has on function of Golgi residential enzymes involved in N-glycosylation...
December 2016: Alcoholism, Clinical and Experimental Research
https://www.readbyqxmd.com/read/27732847/inhibition-of-wave-regulatory-complex-activation-by-a-bacterial-virulence-effector-counteracts-pathogen-phagocytosis
#7
Daniel Humphreys, Vikash Singh, Vassilis Koronakis
To establish pathogenicity, bacteria must evade phagocytosis directed by remodeling of the actin cytoskeleton. We show that macrophages facilitate pathogen phagocytosis through actin polymerization mediated by the WAVE regulatory complex (WRC), small GTPases Arf and Rac1, and the Arf1 activator ARNO. To establish extracellular infections, enteropathogenic (EPEC) and enterohaemorrhagic (EHEC) Escherichia coli hijack the actin cytoskeleton by injecting virulence effectors into the host cell. Here, we find that the virulence effector EspG counteracts WRC-dependent phagocytosis, enabling EPEC and EHEC to remain extracellular...
October 11, 2016: Cell Reports
https://www.readbyqxmd.com/read/27713690/the-endosome-localized-arf-gap-agap1-modulates-dendritic-spine-morphology-downstream-of-the-neurodevelopmental-disorder-factor-dysbindin
#8
Miranda Arnold, Rebecca Cross, Kaela S Singleton, Stephanie Zlatic, Christopher Chapleau, Ariana P Mullin, Isaiah Rolle, Carlene C Moore, Anne Theibert, Lucas Pozzo-Miller, Victor Faundez, Jennifer Larimore
AGAP1 is an Arf1 GTPase activating protein that interacts with the vesicle-associated protein complexes adaptor protein 3 (AP-3) and Biogenesis of Lysosome Related Organelles Complex-1 (BLOC-1). Overexpression of AGAP1 in non-neuronal cells results in an accumulation of endosomal cargoes, which suggests a role in endosome-dependent traffic. In addition, AGAP1 is a candidate susceptibility gene for two neurodevelopmental disorders, autism spectrum disorder (ASD) and schizophrenia (SZ); yet its localization and function in neurons have not been described...
2016: Frontiers in Cellular Neuroscience
https://www.readbyqxmd.com/read/27680705/the-lipolysis-pathway-sustains-normal-and-transformed-stem-cells-in-adult-drosophila
#9
Shree Ram Singh, Xiankun Zeng, Jiangsha Zhao, Ying Liu, Gerald Hou, Hanhan Liu, Steven X Hou
Cancer stem cells (CSCs) may be responsible for tumour dormancy, relapse and the eventual death of most cancer patients. In addition, these cells are usually resistant to cytotoxic conditions. However, very little is known about the biology behind this resistance to therapeutics. Here we investigated stem-cell death in the digestive system of adult Drosophila melanogaster. We found that knockdown of the coat protein complex I (COPI)-Arf79F (also known as Arf1) complex selectively killed normal and transformed stem cells through necrosis, by attenuating the lipolysis pathway, but spared differentiated cells...
September 28, 2016: Nature
https://www.readbyqxmd.com/read/27632000/analysis-of-arf1-gtpase-dependent-membrane-binding-and-remodeling-using-the-exomer-secretory-vesicle-cargo-adaptor
#10
Jon E Paczkowski, J Christopher Fromme
Protein-protein and protein-membrane interactions play a critical role in shaping biological membranes through direct physical contact with the membrane surface. This is particularly evident in many steps of membrane trafficking, in which proteins deform the membrane and induce fission to form transport carriers. The small GTPase Arf1 and related proteins have the ability to remodel membranes by insertion of an amphipathic helix into the membrane. Arf1 and the exomer cargo adaptor coordinate cargo sorting into subset of secretory vesicle carriers in the model organism Saccharomyces cerevisiae...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27582049/assessment-of-housekeeping-genes-as-internal-references-in-quantitative-expression-analysis-during-early-development-of-oyster
#11
Pin Huan, Hongxia Wang, Baozhong Liu
The early development of mollusks exhibits important characteristics from the developmental and evolutionary perspective. With the increasing number of genome-wide studies, accurate analyses of quantitative gene expression during development are impeded by the lack of validated reference genes. To improve the situation, in this study, we analyzed the expression stability of seven candidate housekeeping genes during early development of the Pacific oyster Crassostrea gigas: actin, glyceraldehyde-3 phosphate-dehydrogenase (gapdh), α subunit of elongation factor 1 (elf1α), adp-ribosylation factor 1 (arf1), heterogeneous nuclear ribonucleoprotein q, ubiquitin-conjugating enzyme e2d2 and ribosomal protein s18...
August 31, 2016: Genes & Genetic Systems
https://www.readbyqxmd.com/read/27562622/arf1-and-arf6-regulate-recycling-of-grasp-tamalin-and-the-rac1-gef-dock180-during-hgf-induced-rac1-activation
#12
Emily J Koubek, Lorraine C Santy
Hepatocyte growth factor (HGF) is a potent signaling factor that acts on epithelial cells, causing them to dissociate and scatter. This migration is coordinated by a number of small GTPases, such as ARF6 and Rac1. Active ARF6 is required for HGF-stimulated migration and intracellular levels of ARF6-GTP and Rac1-GTP increase following HGF treatment. During migration, cross talk between ARF6 and Rac1 occurs through formation of a multi-protein complex containing the ARF-GEF cytohesin-2, the scaffolding protein GRASP/Tamalin, and the Rac1-GEF Dock180...
August 12, 2016: Small GTPases
https://www.readbyqxmd.com/read/27535433/the-arf-gap-asap-promotes-arf1-function-at-the-golgi-for-cleavage-furrow-biosynthesis-in-drosophila
#13
Francisco F Rodrigues, Wei Shao, Tony J C Harris
Biosynthetic traffic from the Golgi drives plasma membrane growth. For Drosophila embryo cleavage, this growth is rapid but regulated for cycles of furrow ingression and regression. The highly conserved small G protein Arf1 organizes Golgi trafficking. Arf1 is activated by guanine nucleotide exchange factors, but essential roles for Arf1 GTPase-activating proteins (GAPs) are less clear. We report that the conserved Arf GAP Asap is required for cleavage furrow ingression in the early embryo. Because Asap can affect multiple subcellular processes, we used genetic approaches to dissect its primary effect...
October 15, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27517156/suppression-of-breast-cancer-metastasis-through-the-inactivation-of-adp-ribosylation-factor-1
#14
Xiayang Xie, Shou-Ching Tang, Yafei Cai, Wenhu Pi, Libin Deng, Guangyu Wu, Alain Chavanieu, Yong Teng
Metastasis is the major cause of cancer-related death in breast cancer patients, which is controlled by specific sets of genes. Targeting these genes may provide a means to delay cancer progression and allow local treatment to be more effective. We report for the first time that ADP-ribosylation factor 1 (ARF1) is the most amplified gene in ARF gene family in breast cancer, and high-level amplification of ARF1 is associated with increased mRNA expression and poor outcomes of patients with breast cancer. Knockdown of ARF1 leads to significant suppression of migration and invasion in breast cancer cells...
August 10, 2016: Oncotarget
https://www.readbyqxmd.com/read/27500526/spatial-temporal-study-of-rab1b-dynamics-and-function-at-the-er-golgi-interface
#15
Hernán Martinez, Iris A García, Luciana Sampieri, Cecilia Alvarez
The GTPase Rab1b is involved in ER to Golgi transport, with multiple Rab1b effectors (located at ERES, VTCs and the Golgi complex) being required for its function. In this study, we performed live-cell dual-expression studies to analyze the dynamics of Rab1b and some effectors located at the ERES-Golgi interface. Rab1b occupied widely distributed mobile punctate and tubular structures, displaying a transient overlaps with its effectors and showing that these overlaps occurred at the same time in spatially distinct steps of ER to Golgi transport...
2016: PloS One
https://www.readbyqxmd.com/read/27420958/expressed-sequence-tag-analysis-of-functional-genes-associated-with-adventitious-rooting-in-liriodendron-hybrids
#16
Y D Zhong, X Y Sun, E Y Liu, Y Q Li, Z Gao, F X Yu
Liriodendron hybrids (Liriodendron chinense x L. tulipifera) are important landscaping and afforestation hardwood trees. To date, little genomic research on adventitious rooting has been reported in these hybrids, as well as in the genus Liriodendron. In the present study, we used adventitious roots to construct the first cDNA library for Liriodendron hybrids. A total of 5176 expressed sequence tags (ESTs) were generated and clustered into 2921 unigenes. Among these unigenes, 2547 had significant homology to the non-redundant protein database representing a wide variety of putative functions...
2016: Genetics and Molecular Research: GMR
https://www.readbyqxmd.com/read/27373159/structural-insights-into-arl1-mediated-targeting-of-the-arf-gef-big1-to-the-trans-golgi
#17
Antonio Galindo, Nicolas Soler, Stephen H McLaughlin, Minmin Yu, Roger L Williams, Sean Munro
The GTPase Arf1 is the major regulator of vesicle traffic at both the cis- and trans-Golgi. Arf1 is activated at the cis-Golgi by the guanine nucleotide exchange factor (GEF) GBF1 and at the trans-Golgi by the related GEF BIG1 or its paralog, BIG2. The trans-Golgi-specific targeting of BIG1 and BIG2 depends on the Arf-like GTPase Arl1. We find that Arl1 binds to the dimerization and cyclophilin binding (DCB) domain in BIG1 and report a crystal structure of human Arl1 bound to this domain. Residues in the DCB domain that bind Arl1 are required for BIG1 to locate to the Golgi in vivo...
July 19, 2016: Cell Reports
https://www.readbyqxmd.com/read/27344941/activation-of-aplysia-arf6-induces-neurite-outgrowth-and-is-sequestered-by-the-overexpression-of-the-ph-domain-of-aplysia-sec7-proteins
#18
Deok-Jin Jang, Yong-Woo Jun, Jaehoon Shim, Su-Eon Sim, Jin-A Lee, Chae-Seok Lim, Bong-Kiun Kaang
ADP-ribosylation factors (ARFs) are small guanosine triphosphatases of the Ras superfamily involved in membrane trafficking and regulation of the actin cytoskeleton. Aplysia Sec7 protein (ApSec7), a guanine nucleotide exchange factor for ARF1 and ARF6, induces neurite outgrowth and plays a key role in 5-hydroxyltryptamine-induced neurite growth and synaptic facilitation in Aplysia sensory-motor synapses. However, the specific role of ARF6 signaling on neurite outgrowth in Aplysia neurons has not been examined...
June 21, 2016: Neurobiology of Learning and Memory
https://www.readbyqxmd.com/read/27335351/n-glycomic-and-microscopic-subcellular-localization-analyses-of-npp1-2-and-6-strongly-indicate-that-trans-golgi-compartments-participate-in-the-golgi-to-plastid-traffic-of-nucleotide-pyrophosphatase-phosphodiesterases-in-rice
#19
Kentaro Kaneko, Takeshi Takamatsu, Takuya Inomata, Kazusato Oikawa, Kimiko Itoh, Kazuko Hirose, Maho Amano, Shin-Ichiro Nishimura, Kiminori Toyooka, Ken Matsuoka, Javier Pozueta-Romero, Toshiaki Mitsui
Nucleotide pyrophosphatase/phosphodiesterases (NPPs) are widely distributed N-glycosylated enzymes that catalyze the hydrolytic breakdown of numerous nucleotides and nucleotide sugars. In many plant species, NPPs are encoded by a small multigene family, which in rice are referred to NPP1-NPP6 Although recent investigations showed that N-glycosylated NPP1 is transported from the endoplasmic reticulum (ER)-Golgi system to the chloroplast through the secretory pathway in rice cells, information on N-glycan composition and subcellular localization of other NPPs is still lacking...
August 2016: Plant & Cell Physiology
https://www.readbyqxmd.com/read/27320911/cholesterol-independent-srebp-1-maturation-is-linked-to-arf1-inactivation
#20
Lorissa J Smulan, Wei Ding, Elizaveta Freinkman, Sharvari Gujja, Yvonne J K Edwards, Amy K Walker
Lipogenesis requires coordinated expression of genes for fatty acid, phospholipid, and triglyceride synthesis. Transcription factors, such as SREBP-1 (Sterol regulatory element binding protein), may be activated in response to feedback mechanisms linking gene activation to levels of metabolites in the pathways. SREBPs can be regulated in response to membrane cholesterol and we also found that low levels of phosphatidylcholine (a methylated phospholipid) led to SBP-1/SREBP-1 maturation in C. elegans or mammalian models...
June 28, 2016: Cell Reports
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