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https://www.readbyqxmd.com/read/28930671/cohesin-can-remain-associated-with-chromosomes-during-dna-replication
#1
James D P Rhodes, Judith H I Haarhuis, Jonathan B Grimm, Benjamin D Rowland, Luke D Lavis, Kim A Nasmyth
To ensure disjunction to opposite poles during anaphase, sister chromatids must be held together following DNA replication. This is mediated by cohesin, which is thought to entrap sister DNAs inside a tripartite ring composed of its Smc and kleisin (Scc1) subunits. How such structures are created during S phase is poorly understood, in particular whether they are derived from complexes that had entrapped DNAs prior to replication. To address this, we used selective photobleaching to determine whether cohesin associated with chromatin in G1 persists in situ after replication...
September 19, 2017: Cell Reports
https://www.readbyqxmd.com/read/28924046/in-silico-modeling-of-the-cryptic-e2-ubiquitin-binding-site-of-e6-associated-protein-e6ap-ube3a-reveals-the-mechanism-of-polyubiquitin-chain-assembly
#2
Virginia P Ronchi, Elizabeth D Kim, Christopher M Summa, Jennifer M Klein, Arthur L Haas
To understand the mechanism for assembly of Lys(48)-linked polyubiquitin degradation signals, we previously demonstrated the E6AP/UBE3A ligase harbors two functionally-distinct E2~ubiquitin binding sites: a high-affinity Site 1 required for E6AP Cys(820)~ubiquitin thioester formation and a canonical Site 2 responsible for subsequent chain elongation. Ordered binding to Sites 1 and 2 is here revealed by observation of UbcH7~ubiquitin-dependent substrate inhibition of chain formation at micromolar concentrations...
September 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28914091/short-term-carbon-dynamics-in-a-temperate-grassland-and-heathland-ecosystem-exposed-to-104-days-of-drought-followed-by-irrigation
#3
Kavita Srivastava, Anke Jentsch, Juergen Kreyling, Bruno Glaser, Guido L B Wiesenberg
Temperate ecosystems are susceptible to drought events. The effect of a severe drought (104 days) followed by irrigation on the plant C uptake, its assimilation and input of C in soil were examined using a triple (13)CO2 pulse-chase labelling experiment in model grassland and heathland ecosystems. First (13)CO2 pulse at day 0 of the experiment revealed much higher (13)C tracer uptake for shoots, roots and soil compared to the second pulse (day 44), where all plants showed significantly lower (13)C tracer uptake...
September 15, 2017: Isotopes in Environmental and Health Studies
https://www.readbyqxmd.com/read/28900007/e46k-%C3%AE-synuclein-pathological-mutation-causes-cell-autonomous-toxicity-without-altering-protein-turnover-or-aggregation
#4
Ignacio Íñigo-Marco, Miguel Valencia, Laura Larrea, Ricardo Bugallo, Mikel Martínez-Goikoetxea, Iker Zuriguel, Montserrat Arrasate
α-Synuclein (aSyn) is the main driver of neurodegenerative diseases known as "synucleinopathies," but the mechanisms underlying this toxicity remain poorly understood. To investigate aSyn toxic mechanisms, we have developed a primary neuronal model in which a longitudinal survival analysis can be performed by following the overexpression of fluorescently tagged WT or pathologically mutant aSyn constructs. Most aSyn mutations linked to neurodegenerative disease hindered neuronal survival in this model; of these mutations, the E46K mutation proved to be the most toxic...
September 12, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28860962/cell-proliferation-migration-and-neurogenesis-in-the-adult-brain-of-the-pulse-type-weakly-electric-fish-gymnotus-omarorum
#5
Valentina Olivera-Pasilio, Moira Lasserre, María E Castelló
Adult neurogenesis, an essential mechanism of brain plasticity, enables brain development along postnatal life, constant addition of new neurons, neuronal turnover, and/or regeneration. It is amply distributed but negatively modulated during development and along evolution. Widespread cell proliferation, high neurogenic, and regenerative capacities are considered characteristics of teleost brains during adulthood. These anamniotes are promising models to depict factors that modulate cell proliferation, migration, and neurogenesis, and might be intervened to promote brain plasticity in mammals...
2017: Frontiers in Neuroscience
https://www.readbyqxmd.com/read/28860004/lox-1-a-potential-target-for-therapy-in-atherosclerosis-an-in-vitro-study
#6
Albina Arjuman, Nimai C Chandra
Pro-inflammatory signal generated from the interaction of oxLDL with its cognate receptor LOX-1 has been attenuated successfully by a novel combination siRNA (siLOX-1(Ω)) targeting unique regions of Homo sapien LOX-1 mRNA. Signalling via LOX-1R was studied in a potentially pro-atherogenic arena recreated in a metabolic, pulse-chase set up. An initial pulse of oxLDL (20μg/mL;5h) was chased (without oxLDL) on a temporal scale upto 72h. Our study shows that the pro-inflammatory signal generated via oxLDL-LOX-1R interaction was mediated in two rungs, an initial sustained increase in LOX-1R expression up to 12h, and a renewal after 48h...
August 30, 2017: International Journal of Biochemistry & Cell Biology
https://www.readbyqxmd.com/read/28851751/mrna-pseudouridylation-affects-rna-metabolism-in-the-parasite-toxoplasma-gondii
#7
Margaret A Nakamoto, Alexander F Lovejoy, Alicja M Cygan, John C Boothroyd
RNA contains over 100 modified nucleotides that are created post-transcriptionally, among which pseudouridine (Ψ) is one of the most abundant. Although it was one of the first modifications discovered, the biological role of this modification is still not fully understood. Recently, we reported that a pseudouridine synthase (TgPUS1) is necessary for differentiation of the single-celled eukaryotic parasite Toxoplasma gondii from active to chronic infection. To better understand the biological role of pseudouridylation we report here gel-based and deep-sequencing methods to identify TgPUS1-dependent Ψs in Toxoplasma RNA, and the use of TgPUS1 mutants to examine the effect of this modification on mRNAs...
August 29, 2017: RNA
https://www.readbyqxmd.com/read/28837266/covalent-dimerization-of-interleukin-like-epithelial-to-mesenchymal-transition-emt-inducer-ilei-facilitates-emt-invasion-and-late-aspects-of-metastasis
#8
Maria Kral, Christoph Klimek, Betül Kutay, Gerald Timelthaler, Thomas Lendl, Benjamin Neuditschko, Christopher Gerner, Maria Sibilia, Agnes Csiszar
The interleukin-like epithelial-to-mesenchymal transition (EMT) inducer (ILEI)/FAM3C is a member of the highly homologous FAM3 family and is essential for EMT and metastasis formation. It is upregulated in several cancers, and its altered subcellular localization strongly correlates with poor survival. However, the mechanism of ILEI action, including the structural requirements for ILEI activity, remains elusive. Here, we show that ILEI formed both monomers and covalent dimers in cancer cell lines and in tumors...
August 24, 2017: FEBS Journal
https://www.readbyqxmd.com/read/28813167/lipophagy-maintains-energy-homeostasis-in-the-kidney-proximal-tubule-during-prolonged-starvation
#9
Satoshi Minami, Takeshi Yamamoto, Yoshitsugu Takabatake, Atsushi Takahashi, Tomoko Namba, Jun Matsuda, Tomonori Kimura, Jun-Ya Kaimori, Isao Matsui, Takayuki Hamano, Hiroaki Takeda, Masatomo Takahashi, Yoshihiro Izumi, Takeshi Bamba, Taiji Matsusaka, Fumio Niimura, Yoshitaka Isaka
Macroautophagy/autophagy is a self-degradation process that combats starvation. Lipids are the main energy source in kidney proximal tubular cells (PTCs). During starvation, PTCs increase fatty acid (FA) uptake, form intracellular lipid droplets (LDs), and hydrolyze them for use. The involvement of autophagy in lipid metabolism in the kidney remains largely unknown. Here, we investigated the autophagy-mediated regulation of renal lipid metabolism during prolonged starvation using PTC-specific Atg5-deficient (atg5-TSKO) mice and an in vitro serum starvation model...
August 16, 2017: Autophagy
https://www.readbyqxmd.com/read/28795080/electron-lattice-energy-relaxation-in-laser-excited-thin-film-au-insulator-heterostructures-studied-by-ultrafast-mev-electron-diffraction
#10
K Sokolowski-Tinten, X Shen, Q Zheng, T Chase, R Coffee, M Jerman, R K Li, M Ligges, I Makasyuk, M Mo, A H Reid, B Rethfeld, T Vecchione, S P Weathersby, H A Dürr, X J Wang
We apply time-resolved MeV electron diffraction to study the electron-lattice energy relaxation in thin film Au-insulator heterostructures. Through precise measurements of the transient Debye-Waller-factor, the mean-square atomic displacement is directly determined, which allows to quantitatively follow the temporal evolution of the lattice temperature after short pulse laser excitation. Data obtained over an extended range of laser fluences reveal an increased relaxation rate when the film thickness is reduced or the Au-film is capped with an additional insulator top-layer...
September 2017: Structural Dynamics (Melville, N.Y.)
https://www.readbyqxmd.com/read/28760965/quantitative-time-resolved-chemoproteomics-reveals-that-stable-o-glcnac-regulates-box-c-d-snornp-biogenesis
#11
Wei Qin, Pinou Lv, Xinqi Fan, Baiyi Quan, Yuntao Zhu, Ke Qin, Ying Chen, Chu Wang, Xing Chen
O-linked GlcNAcylation (O-GlcNAcylation), a ubiquitous posttranslational modification on intracellular proteins, is dynamically regulated in cells. To analyze the turnover dynamics of O-GlcNAcylated proteins, we developed a quantitative time-resolved O-linked GlcNAc proteomics (qTOP) strategy based on metabolic pulse-chase labeling with an O-GlcNAc chemical reporter and stable isotope labeling with amino acids in cell culture (SILAC). Applying qTOP, we quantified the turnover rates of 533 O-GlcNAcylated proteins in NIH 3T3 cells and discovered that about 14% exhibited minimal removal of O-GlcNAc or degradation of protein backbones...
August 15, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28760849/stability-of-proteins-out-of-service-the-gapb-case-of-bacillus-subtilis
#12
Ulf Gerth, Eleonora Krieger, Daniela Zühlke, Alexander Reder, Uwe Völker, Michael Hecker
Bacillus subtilis possesses two glyceraldehyde-3-phosphate dehydrogenases with opposite roles, the glycolytic NAD-dependent GapA and the NADP-dependent GapB enzyme, which is exclusively required during gluconeogenesis but not active under conditions promoting glycolysis. We propose, that proteins no longer needed will be recognized and proteolyzed by Clp proteases and thereby recycled. To test this postulation, we analyzed the stability of the glycolytic enzyme GapA and the gluconeogenetic enzyme GapB in the presence and absence of glucose...
July 31, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/28751886/salmonella-enterica-serovar-infantis-from-food-and-human-infections-switzerland-2010-2015-poultry-related-multidrug-resistant-clones-and-an-emerging-esbl-producing-clonal-lineage
#13
Denise Hindermann, Gopal Gopinath, Hannah Chase, Flavia Negrete, Denise Althaus, Katrin Zurfluh, Ben D Tall, Roger Stephan, Magdalena Nüesch-Inderbinen
Objectives: The aim of this study was to characterize a collection of 520 Salmonella enterica serovar Infantis strains isolated from food (poultry meat), human infections and environmental sources from the years 2010, 2013 and 2015 in Switzerland. Methods: We performed antimicrobial susceptibility testing and pulsed-field gel electrophoresis (PFGE) analysis on all 520 S. Infantis isolates, and whole genome sequencing (WGS) on 32 selected isolates. Results: The majority (74.8%) of the isolates was multidrug resistant (MDR)...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28720784/snap-tagged-chikungunya-virus-replicons-improve-visualisation-of-non-structural-protein-3-by-fluorescence-microscopy
#14
Roland Remenyi, Grace C Roberts, Carsten Zothner, Andres Merits, Mark Harris
Chikungunya virus (CHIKV), a mosquito-borne alphavirus, causes febrile disease, muscle and joint pain, which can become chronic in some individuals. The non-structural protein 3 (nsP3) plays essential roles during infection, but a complete understanding of its function is lacking. Here we used a microscopy-based approach to image CHIKV nsP3 inside human cells. The SNAP system consists of a self-labelling enzyme tag, which catalyses the covalent linking of exogenously supplemented synthetic ligands. Genetic insertion of this tag resulted in viable replicons and specific labelling while preserving the effect of nsP3 on stress granule responses and co-localisation with GTPase Activating Protein (SH3 domain) Binding Proteins (G3BPs)...
July 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28717648/response-detection-of-castrate-resistant-prostate-cancer-to-clinically-utilised-and-novel-treatments-by-monitoring-phospholipid-metabolism
#15
Tim A D Smith, Su M Phyu, Kholoud S Alzyoud, Chih-Chung Tseng
Androgen receptor (AR) activation is the primary driving factor in prostate cancer which is initially responsive to castration but then becomes resistant (castration-resistant prostate cancer (CRPC)). CRPC cells still retain the functioning AR which can be targeted by other therapies. A recent promising development is the use of inhibitors (Epi-1) of protein-protein interaction to inhibit AR-activated signalling. Translating novel therapies into the clinic requires sensitive early response indicators. Here potential response markers are explored...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28710114/circadian-rhythm-disruption-impairs-tissue-homeostasis-and-exacerbates-chronic-inflammation-in-the-intestine
#16
René Pagel, Florian Bär, Torsten Schröder, Annika Sünderhauf, Axel Künstner, Saleh M Ibrahim, Stella E Autenrieth, Kathrin Kalies, Peter König, Anthony H Tsang, Dominik Bettenworth, Senad Divanovic, Hendrik Lehnert, Klaus Fellermann, Henrik Oster, Stefanie Derer, Christian Sina
Endogenous circadian clocks regulate 24 h rhythms of physiology and behavior. Circadian rhythm disruption (CRD) is suggested as a risk factor for inflammatory bowel disease. However, the underlying molecular mechanisms remain unknown. Intestinal biopsies from Per1/2 mutant and wild-type (WT) mice were investigated by electron microscopy, immunohistochemistry, and bromodeoxyuridine pulse-chase experiments. TNF-α was injected intraperitoneally, with or without necrostatin-1, into Per1/2 mice or rhythmic and externally desynchronized WT mice to study intestinal epithelial cell death...
July 14, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28691764/cten-promotes-epithelial-mesenchymal-transition-through-the-post-transcriptional-stabilization-of-snail
#17
Hannah Thorpe, Abdulaziz Asiri, Maham Akhlaq, Mohammad Ilyas
Cten promotes cell migration however the knowledge of underlying signalling pathways is sparse. We have shown that Cten downregulates E-cadherin, a feature of epithelial to mesenchymal transition (EMT). This prompted us to investigate whether Cten further contributed to EMT processes to regulate cell motility. The regulation of Snail by Cten was investigated following overexpression, knockdown (by RNA-interference) or knockout of Cten in HCT116, Caco-2 and SW620 colorectal cancer (CRC) cell lines. Subsequently, the cycloheximide (CHX) pulse chase assay was used to investigate changes in Snail protein stability and the functional relevance of Cten-Snail signalling was investigated...
July 10, 2017: Molecular Carcinogenesis
https://www.readbyqxmd.com/read/28686945/trophic-transfer-of-134-cs-in-the-manila-clam-ruditapes-philippinarum
#18
Murat Belivermiş, Önder Kılıç, Narin Sezer, Gülşah Kalaycı, Marc Metian
Bioaccumulation of radiocaesium in many marine organisms occurs through complex trophic transfer mechanisms. The present study addresses the trophic transfer of (134)Cs in the widely distributed marine bivalve, the Manila clam Ruditapes philippinarum, by experimentally determining the assimilation efficiency (AE) and the specific role of food quality or diet on the AE in this marine invertebrate. Pulse-chase feeding experiments were carried out on this clam using the phytoplankton species Tetraselmis chuii, Phaeodactylum tricornutum and Isochrysis galbana...
October 2017: Journal of Environmental Radioactivity
https://www.readbyqxmd.com/read/28684414/kinetic-processivity-of-the-two-step-oxidations-of-progesterone-and-pregnenolone-to-androgens-by-human-cytochrome-p450-17a1
#19
COMPARATIVE STUDY
Eric Gonzalez, F Peter Guengerich
Cytochrome P450 (P450, CYP) 17A1 plays a critical role in steroid metabolism, catalyzing both the 17α-hydroxylation of pregnenolone and progesterone and the subsequent 17α,20-lyase reactions to form dehydroepiandrosterone (DHEA) and androstenedione (Andro), respectively, critical for generating glucocorticoids and androgens. Human P450 17A1 reaction rates examined are enhanced by the accessory protein cytochrome b5 (b5), but the exact role of b5 in P450 17A1-catalyzed reactions is unclear as are several details of these reactions...
August 11, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28650345/preexisting-endothelial-cells-mediate-cardiac-neovascularization-after-injury
#20
Lingjuan He, Xiuzhen Huang, Onur Kanisicak, Yi Li, Yue Wang, Yan Li, Wenjuan Pu, Qiaozhen Liu, Hui Zhang, Xueying Tian, Huan Zhao, Xiuxiu Liu, Shaohua Zhang, Yu Nie, Shengshou Hu, Xiang Miao, Qing-Dong Wang, Fengchao Wang, Ting Chen, Qingbo Xu, Kathy O Lui, Jeffery D Molkentin, Bin Zhou
The mechanisms that promote the generation of new coronary vasculature during cardiac homeostasis and after injury remain a fundamental and clinically important area of study in the cardiovascular field. Recently, it was reported that mesenchymal-to-endothelial transition (MEndoT) contributes to substantial numbers of coronary endothelial cells after myocardial infarction. Therefore, the MEndoT has been proposed as a paradigm mediating neovascularization and is considered a promising therapeutic target in cardiac regeneration...
August 1, 2017: Journal of Clinical Investigation
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