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https://www.readbyqxmd.com/read/27811356/spatio-temporal-regulation-of-egfr-signaling-by-the-eps15-homology-domain-containing-protein-3-ehd3
#1
Mohamed Amessou, Abdul Shukkur Ebrahim, Ashok Dilly, Melvin Joseph, Marina Tabolina, Sahiti Chukkapalli, Louay Meroueh, Joseph T Syed, Allison Liddane, Siera Lanae Lang, Ayad Al-Katib, Mustapha Kandouz
The epidermal growth factor (EGF) receptor EGFR is a major receptor tyrosine kinase whose role in gliomagenesis is well established. We have recently identified EHD3 [Eps15 homology (EH) domain-containing protein 3], an endocytic trafficking regulatory protein, as a putative brain tumor suppressor. Here, we investigate the underlying mechanisms, by establishing a novel mechanistic and functional connection between EHD3 and the EGFR signaling pathway. We show that, in response to stimulation with the EGF ligand, EHD3 accelerates the rate of EGFR degradation by dramatically increasing its ubiquitination...
November 1, 2016: Oncotarget
https://www.readbyqxmd.com/read/27670116/npf-motifs-in-the-vaccinia-virus-protein-a36-recruit-intersectin-1-to-promote-cdc42-n-wasp-mediated-viral-release-from-infected-cells
#2
Xenia Snetkov, Ina Weisswange, Julia Pfanzelter, Ashley C Humphries, Michael Way
During its egress, vaccinia virus transiently recruits AP-2 and clathrin after fusion with the plasma membrane. This recruitment polarizes the viral protein A36 beneath the virus, enhancing actin polymerization and the spread of infection. We now demonstrate that three NPF motifs in the C-terminus of A36 recruit AP-2 and clathrin by interacting directly with the Epsin15 homology domains of Eps15 and intersectin-1. A36 is the first identified viral NPF motif containing protein shown to interact with endocytic machinery...
2016: Nature Microbiology
https://www.readbyqxmd.com/read/27611902/analysis-of-loops-that-mediate-protein-protein-interactions-and-translation-into-submicromolar-inhibitors
#3
Timothy R Siegert, Michael J Bird, Kamlesh M Makwana, Joshua A Kritzer
Effective strategies for mimicking α-helix and β-strand epitopes have been developed, producing valuable inhibitors for some classes of protein-protein interactions (PPIs). However, there are no general strategies for translating loop epitopes into useful PPI inhibitors. In this work, we use the LoopFinder program to identify diverse sets of "hot loops," which are loop epitopes that mediate PPIs. These include loops that are well-suited to mimicry with macrocyclic compounds, and loops that are most similar to variable loops on antibodies and ankyrin repeat proteins...
October 5, 2016: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/27557559/epsin1-modulates-synaptic-vesicle-retrieval-capacity-at-cns-synapses
#4
Jae Won Kyung, Jae Ryul Bae, Dae-Hwan Kim, Woo Keun Song, Sung Hyun Kim
Synaptic vesicle retrieval is an essential process for continuous maintenance of neural information flow after synaptic transmission. Epsin1, originally identified as an EPS15-interacting protein, is a major component of clathrin-mediated endocytosis. However, the role of Epsin1 in synaptic vesicle endocytosis at CNS synapses remains elusive. Here, we showed significantly altered synaptic vesicle endocytosis in neurons transfected with shRNA targeting Epsin1 during/after neural activity. Endocytosis was effectively restored by introducing shRNA-insensitive Epsin1 into Epsin1-depleted neurons...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27531895/mammalian-eps15-homology-domain-1-promotes-metastasis-in-non-small-cell-lung-cancer-by-inducing-epithelial-mesenchymal-transition
#5
Qingwei Meng, Ying Xing, Tingting Ren, Hailing Lu, Yuhui Xi, Zhijun Jiang, Jing Hu, Chunhong Li, Lichun Sun, Dianjun Sun, Li Cai
The identification of the earliest molecular events responsible for the metastatic dissemination of non-small cell lung cancer (NSCLC) remains critical for early detection, prevention, and treatment interventions. In this study, we hypothesized that Mammalian Eps15 homology domain 1 (EHD1) might be responsible for the metastatic behavior of cells in NSCLC. We demonstrated that upregulation of EHD1 is associated with lymph nodes metastasis and unfavorable survival in patients with NSCLC. EHD1 knockdown inhibited the invasion and migration of human NSCLC cells, and overexpression of EHD1 increased the metastatic potential of lung cancer cells...
August 11, 2016: Oncotarget
https://www.readbyqxmd.com/read/27492965/agonist-induced-internalization-and-desensitization-of-the-apelin-receptor
#6
George R Pope, Sharada Tilve, Craig A McArdle, Stephen J Lolait, Anne-Marie O'Carroll
Apelin acts via the G protein-coupled apelin receptor (APJ) to mediate effects on cardiovascular and fluid homeostasis. G protein-coupled receptor (GPCR) trafficking has an important role in the regulation of receptor signalling pathways and cellular functions, however in the case of APJ the mechanisms and proteins involved in apelin-induced trafficking are not well understood. We generated a stable HEK-293 cell line expressing N-terminus HA-tagged mouse (m) APJ, and used a semi-automated imaging protocol to quantitate APJ trafficking and ERK1/2 activation following stimulation with [Pyr(1)]apelin-13...
December 5, 2016: Molecular and Cellular Endocrinology
https://www.readbyqxmd.com/read/27489278/entry-of-classical-swine-fever-virus-into-pk-15-cells-via-a-ph-dynamin-and-cholesterol-dependent-clathrin-mediated-endocytic-pathway-that-requires-rab5-and-rab7
#7
Bao-Jun Shi, Chun-Chun Liu, Jing Zhou, Shi-Qi Wang, Zhi-Can Gao, Xiao-Min Zhang, Bin Zhou, Pu-Yan Chen
UNLABELLED: Classical swine fever virus (CSFV), a member of the genus Pestivirus within the family Flaviviridae, is a small, enveloped, positive-strand RNA virus. Due to its economic importance to the pig industry, the biology and pathogenesis of CSFV have been investigated extensively. However, the mechanisms of CSFV entry into cells are not well characterized. In this study, we used systematic approaches to dissect CSFV cell entry. We first observed that CSFV infection was inhibited by chloroquine and NH4Cl, suggesting that viral entry required a low-pH environment...
October 15, 2016: Journal of Virology
https://www.readbyqxmd.com/read/27411790/ehd1-confers-resistance-to-cisplatin-in-non-small-cell-lung-cancer-by-regulating-intracellular-cisplatin-concentrations
#8
Jing Gao, Qingwei Meng, Yanbin Zhao, Xuesong Chen, Li Cai
BACKGROUND: Non-small cell lung cancer (NSCLC) is one of the most aggressive types of cancer. However, resistance to cisplatin (CDDP) remains a major challenge in NSCLC treatment. The purpose of this study was to investigate the ability of EHD1 [Eps15 homology (EH) domain - containing protein 1] to confer CDDP resistance in NSCLC cells and to investigate mechanisms of this resistance. METHODS: The associations between EHD1 expression in NSCLC specimens and clinicopathological features, including prognosis, were assessed by immunohistochemistry (IHC)...
2016: BMC Cancer
https://www.readbyqxmd.com/read/27385443/how-antibodies-alter-the-cell-entry-pathway-of-dengue-virus-particles-in-macrophages
#9
Nilda V Ayala-Nunez, Tabitha E Hoornweg, Denise P I van de Pol, Klaas A Sjollema, Jacky Flipse, Hilde M van der Schaar, Jolanda M Smit
Antibody-dependent enhancement of dengue virus (DENV) infection plays an important role in the exacerbation of DENV-induced disease. To understand how antibodies influence the fate of DENV particles, we explored the cell entry pathway of DENV in the absence and presence of antibodies in macrophage-like P388D1 cells. Recent studies unraveled that both mature and immature DENV particles contribute to ADE, hence, both particles were studied. We observed that antibody-opsonized DENV enters P388D1 cells through a different pathway than non-opsonized DENV...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27385343/eps15-membrane-binding-and-bending-activity-acts-redundantly-with-fcho1-during-clathrin-mediated-endocytosis
#10
Lei Wang, Adam Johnson, Michael Hanna, Anjon Audhya
Clathrin coat assembly on membranes requires cytosolic adaptors and accessory proteins, which bridge triskeleons with the lipid bilayer and stabilize lattice architecture throughout the process of vesicle formation. In Caenorhabditis elegans, the prototypical AP-2 adaptor complex, which is activated by the accessory factor Fcho1 at the plasma membrane, is dispensable during embryogenesis, enabling us to define alternative mechanisms that facilitate clathrin-mediated endocytosis. Here we uncover a synthetic genetic interaction between C...
September 1, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27237791/transient-fcho1-2%C3%A2-eps15-r%C3%A2-ap-2-nanoclusters-prime-the-ap-2-clathrin-adaptor-for-cargo-binding
#11
Li Ma, Perunthottathu K Umasankar, Antoni G Wrobel, Anastasia Lymar, Airlie J McCoy, Sachin S Holkar, Anupma Jha, Tirthadipa Pradhan-Sundd, Simon C Watkins, David J Owen, Linton M Traub
Clathrin-coated vesicles form by rapid assembly of discrete coat constituents into a cargo-sorting lattice. How the sequential phases of coat construction are choreographed is unclear, but transient protein-protein interactions mediated by short interaction motifs are pivotal. We show that arrayed Asp-Pro-Phe (DPF) motifs within the early-arriving endocytic pioneers Eps15/R are differentially decoded by other endocytic pioneers Fcho1/2 and AP-2. The structure of an Eps15/R⋅Fcho1 μ-homology domain complex reveals a spacing-dependent DPF triad, bound in a mechanistically distinct way from the mode of single DPF binding to AP-2...
June 6, 2016: Developmental Cell
https://www.readbyqxmd.com/read/27224507/csf-1-receptor-signalling-is-governed-by-pre-requisite-ehd1-mediated-receptor-display-on-the-macrophage-cell-surface
#12
Luke R Cypher, Timothy Alan Bielecki, Lu Huang, Wei An, Fany Iseka, Eric Tom, Matthew D Storck, Adam D Hoppe, Vimla Band, Hamid Band
Colony stimulating factor-1 receptor (CSF-1R), a receptor tyrosine kinase (RTK), is the master regulator of macrophage biology. CSF-1 can bind CSF-1R resulting in receptor activation and signalling essential for macrophage functions such as proliferation, differentiation, survival, polarization, phagocytosis, cytokine secretion, and motility. CSF-1R activation can only occur after the receptor is presented on the macrophage cell surface. This process is reliant upon the underlying macrophage receptor trafficking machinery...
September 2016: Cellular Signalling
https://www.readbyqxmd.com/read/27221498/decreased-expression-of-ehd2-promotes-tumor-metastasis-and-indicates-poor-prognosis-in-hepatocellular-carcinoma
#13
Jinxia Liu, Wenkai Ni, Lishuai Qu, Xiaopeng Cui, Zhipeng Lin, Qingqing Liu, Huiling Zhou, Runzhou Ni
BACKGROUND: Metastasis remains the most common cause of lethal outcomes in hepatocellular carcinoma (HCC) after curative resection. Understanding molecular mechanisms that regulate metastasis process is crucial for improving treatment of hepatocellular carcinoma. AIMS: In this article, we examined whether Eps15 homology domain-containing 2 (EHD2) played a critical role in hepatocellular carcinoma metastasis and explored the possible mechanism. METHODS: EHD2 and E-cadherin expression levels in hepatocellular carcinoma patients were examined using Western blotting and immunohistochemistry...
September 2016: Digestive Diseases and Sciences
https://www.readbyqxmd.com/read/27189942/ehd3-protein-is-required-for-tubular-recycling-endosome-stabilization-and-an-asparagine-glutamic-acid-residue-pair-within-its-eps15-homology-eh-domain-dictates-its-selective-binding-to-npf-peptides
#14
Kriti Bahl, Shuwei Xie, Gaelle Spagnol, Paul Sorgen, Naava Naslavsky, Steve Caplan
An elaborate network of dynamic lipid membranes, termed tubular recycling endosomes (TRE), coordinates the process of endocytic recycling in mammalian cells. The C-terminal Eps15 homology domain (EHD)-containing proteins have been implicated in the bending and fission of TRE, thus regulating endocytic recycling. EHD proteins have an EH domain that interacts with proteins containing an NPF motif. We found that NPF-containing EHD1 interaction partners such as molecules interacting with CasL-like1 (MICAL-L1) and Syndapin2 are essential for TRE biogenesis...
June 24, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27170354/dictyostelium-ehd-associates-with-dynamin-and-participates-in-phagosome-maturation
#15
Aurélie Gueho, Cristina Bosmani, Navin Gopaldass, Virginie Molle, Thierry Soldati, François Letourneur
Proteins that contain Eps15 homology domains (EHDs) in their C-terminus are newly identified key regulators of endosomal membrane trafficking. Here, we show that D. discoideum contains a single EHD protein (referred to as EHD) that localizes to endosomal compartments and newly formed phagosomes. We provide the first evidence that EHD regulates phagosome maturation. Deletion of EHD results in defects in intraphagosomal proteolysis and acidification. These defects are linked to early delivery of lysosomal enzymes and fast retrieval of the vacuolar H(+)-ATPase in maturing phagosomes...
June 15, 2016: Journal of Cell Science
https://www.readbyqxmd.com/read/27008177/a-spopl-cullin-3-ubiquitin-ligase-complex-regulates-endocytic-trafficking-by-targeting-eps15-at-endosomes
#16
Michaela Gschweitl, Anna Ulbricht, Christopher A Barnes, Radoslav I Enchev, Ingrid Stoffel-Studer, Nathalie Meyer-Schaller, Jatta Huotari, Yohei Yamauchi, Urs F Greber, Ari Helenius, Matthias Peter
Cullin-3 (CUL3)-based ubiquitin ligases regulate endosome maturation and trafficking of endocytic cargo to lysosomes in mammalian cells. Here, we report that these functions depend on SPOPL, a substrate-specific CUL3 adaptor. We find that SPOPL associates with endosomes and is required for both the formation of multivesicular bodies (MVBs) and the endocytic host cell entry of influenza A virus. In SPOPL-depleted cells, endosomes are enlarged and fail to acquire intraluminal vesicles (ILVs). We identify a critical substrate ubiquitinated by CUL3-SPOPL as EPS15, an endocytic adaptor that also associates with the ESCRT-0 complex members HRS and STAM on endosomes...
2016: ELife
https://www.readbyqxmd.com/read/26914139/yeast-eps15-like-endocytic-protein-pan1p-regulates-the-interaction-between-endocytic-vesicles-endosomes-and-the-actin-cytoskeleton
#17
Junko Y Toshima, Eri Furuya, Makoto Nagano, Chisa Kanno, Yuta Sakamoto, Masashi Ebihara, Daria Elisabeth Siekhaus, Jiro Toshima
The actin cytoskeleton plays important roles in the formation and internalization of endocytic vesicles. In yeast, endocytic vesicles move towards early endosomes along actin cables, however, the molecular machinery regulating interaction between endocytic vesicles and actin cables is poorly understood. The Eps15-like protein Pan1p plays a key role in actin-mediated endocytosis and is negatively regulated by Ark1 and Prk1 kinases. Here we show that pan1 mutated to prevent phosphorylation at all 18 threonines, pan1-18TA, displayed almost the same endocytic defect as ark1Δ prk1Δ cells, and contained abnormal actin concentrations including several endocytic compartments...
2016: ELife
https://www.readbyqxmd.com/read/26884322/endocytic-recycling-protein-ehd1-regulates-primary-cilia-morphogenesis-and-shh-signaling-during-neural-tube-development
#18
Sohinee Bhattacharyya, Mark A Rainey, Priyanka Arya, Samikshan Dutta, Manju George, Matthew D Storck, Rodney D McComb, David Muirhead, Gordon L Todd, Karen Gould, Kaustubh Datta, Janee Gelineau-van Waes, Vimla Band, Hamid Band
Members of the four-member C-terminal EPS15-Homology Domain-containing (EHD) protein family play crucial roles in endocytic recycling of cell surface receptors from endosomes to the plasma membrane. In this study, we show that Ehd1 gene knockout in mice on a predominantly B6 background is embryonic lethal. Ehd1-null embryos die at mid-gestation with a failure to complete key developmental processes including neural tube closure, axial turning and patterning of the neural tube. We found that Ehd1-null embryos display short and stubby cilia on the developing neuroepithelium at embryonic day 9...
2016: Scientific Reports
https://www.readbyqxmd.com/read/26872775/evolutionary-changes-on-the-way-to-clathrin-mediated-endocytosis-in-animals
#19
Mykola Dergai, Anton Iershov, Olga Novokhatska, Serhii Pankivskyi, Alla Rynditch
Endocytic pathways constitute an evolutionarily ancient system that significantly contributed to the eukaryotic cell architecture and to the diversity of cell type-specific functions and signaling cascades, in particular of metazoans. Here we used comparative proteomic studies to analyze the universal internalization route in eukaryotes, clathrin-mediated endocytosis (CME), to address the issues of how this system evolved and what are its specific features. Among 35 proteins crucially required for animal CME, we identified a subset of 22 proteins common to major eukaryotic branches and 13 gradually acquired during evolution...
March 2016: Genome Biology and Evolution
https://www.readbyqxmd.com/read/26841867/structural-studies-of-the-nedd4-ww-domains-and-their-selectivity-for-the-connexin43-cx43-carboxyl-terminus
#20
Gaelle Spagnol, Fabien Kieken, Jennifer L Kopanic, Hanjun Li, Sydney Zach, Kelly L Stauch, Rosslyn Grosely, Paul L Sorgen
Neuronal precursor cell-expressed developmentally down-regulated 4 (Nedd4) was the first ubiquitin protein ligase identified to interact with connexin43 (Cx43), and its suppressed expression results in accumulation of gap junction plaques at the plasma membrane. Nedd4-mediated ubiquitination of Cx43 is required to recruit Eps15 and target Cx43 to the endocytic pathway. Although the Cx43 residues that undergo ubiquitination are still unknown, in this study we address other unresolved questions pertaining to the molecular mechanisms mediating the direct interaction between Nedd4 (WW1-3 domains) and Cx43 (carboxyl terminus (CT))...
April 1, 2016: Journal of Biological Chemistry
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