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https://www.readbyqxmd.com/read/28979270/functional-characterization-of-a-syntaxin-involved-in-tomato-solanum-lycopersicum-resistance-against-powdery-mildew
#1
Valentina Bracuto, Michela Appiano, Zheng Zheng, Anne-Marie A Wolters, Zhe Yan, Luigi Ricciardi, Richard G F Visser, Stefano Pavan, Yuling Bai
Specific syntaxins, such as Arabidopsis AtPEN1 and its barley ortholog ROR2, play a major role in plant defense against powdery mildews. Indeed, the impairment of these genes results in increased fungal penetration in both host and non-host interactions. In this study, a genome-wide survey allowed the identification of 21 tomato syntaxins. Two of them, named SlPEN1a and SlPEN1b, are closely related to AtPEN1. RNAi-based silencing of SlPEN1a in a tomato line carrying a loss-of-function mutation of the susceptibility gene SlMLO1 led to compromised resistance toward the tomato powdery mildew fungus Oidium neolycopersici...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28966893/the-legionella-pneumophila-effector-lpg1137-is-a-homologue-of-mitochondrial-slc25-carrier-proteins-not-of-known-serine-proteases
#2
Marcin Gradowski, Krzysztof Pawłowski
Many bacterial effector proteins that are delivered to host cells during infection are enzymes targeting host cell signalling. Recently, Legionella pneumophila effector Lpg1137 was experimentally characterised as a serine protease that cleaves human syntaxin 17. We present strong bioinformatic evidence that Lpg1137 is a homologue of mitochondrial carrier proteins and is not related to known serine proteases. We also discuss how this finding can be reconciled with the apparently contradictory experimental results...
2017: PeerJ
https://www.readbyqxmd.com/read/28962182/gypenosides-induce-cell-death-and-alter-gene-expression-in-human-oral-cancer-hsc-3-cells
#3
Kung-Wen Lu, Yi-Shih Ma, Fu-Shun Yu, Yi-Ping Huang, Yung-Lin Chu, Rick Sai-Chuen Wu, Ching-Lung Liao, Fu-Shin Chueh, Jing-Gung Chung
Gypenosides (Gyp), the primary components of Gynostemma pentaphyllum Makino, have long been used as a Chinese herbal medicine. In the present study, the effects of Gyp on cell viability, the cell cycle, cell apoptosis, DNA damage and chromatin condensation were investigated in vitro using human oral cancer HSC-3 cells. The results of the present study indicated that Gyp induces cell death, G2/M phase arrest and apoptosis in HSC-3 cells in a dose-dependent manner. It was also demonstrated that Gyp decreased the depolarization of mitochondrial membrane potential in a time-dependent manner...
September 2017: Experimental and Therapeutic Medicine
https://www.readbyqxmd.com/read/28944233/protein-structure-and-phenotypic-analysis-of-pathogenic-and-population-missense-variants-in-stxbp1
#4
Mohnish Suri, Jochem M G Evers, Roman A Laskowski, Sinead O'Brien, Kate Baker, Jill Clayton-Smith, Tabib Dabir, Dragana Josifova, Shelagh Joss, Bronwyn Kerr, Alison Kraus, Meriel McEntagart, Jenny Morton, Audrey Smith, Miranda Splitt, Janet M Thornton, Caroline F Wright
BACKGROUND: Syntaxin-binding protein 1, encoded by STXBP1, is highly expressed in the brain and involved in fusing synaptic vesicles with the plasma membrane. Studies have shown that pathogenic loss-of-function variants in this gene result in various types of epilepsies, mostly beginning early in life. We were interested to model pathogenic missense variants on the protein structure to investigate the mechanism of pathogenicity and genotype-phenotype correlations. METHODS: We report 11 patients with pathogenic de novo mutations in STXBP1 identified in the first 4293 trios of the Deciphering Developmental Disorder (DDD) study, including six missense variants...
September 2017: Molecular Genetics & Genomic Medicine
https://www.readbyqxmd.com/read/28935937/membrane-tension-increases-fusion-efficiency-of-model-membranes-in-the-presence-of-snares
#5
Torben-Tobias Kliesch, Jörn Dietz, Laura Turco, Partho Halder, Elena Polo, Marco Tarantola, Reinhard Jahn, Andreas Janshoff
The large gap in time scales between membrane fusion occurring in biological systems during neurotransmitter release and fusion observed between model membranes has provoked speculations over a large number of possible factors that might explain this discrepancy. One possible reason is an elevated lateral membrane tension present in the presynaptic membrane. We investigated the tension-dependency of fusion using model membranes equipped with a minimal fusion machinery consisting of syntaxin 1, synaptobrevin and SNAP 25...
September 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28933649/legionella-blocks-autophagy-by-cleaving-stx17-syntaxin-17
#6
Kohei Arasaki, Mitsuo Tagaya
Pathogens subvert host defense systems including autophagy and apoptosis for their survival and proliferation. Legionella pneumophila is a Gram-negative bacterium that grows in alveolar macrophages and causes severe pneumonia. Early during infection Legionella secretes effector proteins that convert the plasma membrane-derived vacuole containing Legionella into an endoplasmic reticulum (ER)-like replicative vacuole. These vacuoles ultimately fuse with the ER, where the pathogen replicates. Recently, we showed that one of the effectors, Lpg1137, is a serine protease that targets the mitochondria-associated ER membrane (MAM) and degrades STX17 (syntaxin 17), a SNARE implicated in macroautophagy/autophagy as well as mitochondria dynamics and membrane trafficking in fed cells...
September 21, 2017: Autophagy
https://www.readbyqxmd.com/read/28931085/hcv-induced-autophagosomes-are-generated-via-homotypic-fusion-of-phagophores-that-mediate-hcv-rna-replication
#7
Linya Wang, Ja Yeon Kim, Helene Minyi Liu, Michael M C Lai, Jing-Hsiung James Ou
Hepatitis C virus (HCV) induces autophagy to promote its replication, including its RNA replication, which can take place on double-membrane vesicles known as autophagosomes. However, how HCV induces the biogenesis of autophagosomes and how HCV RNA replication complex may be assembled on autophagosomes were largely unknown. During autophagy, crescent membrane structures known as phagophores first appear in the cytoplasm, which then progress to become autophagosomes. By conducting electron microscopy and in vitro membrane fusion assay, we found that phagophores induced by HCV underwent homotypic fusion to generate autophagosomes in a process dependent on the SNARE protein syntaxin 7 (STX7)...
September 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28914590/canine-salivary-glands-analysis-of-rab-and-snare-protein-expression-and-snare-complex-formation-with-diverse-tissue-properties
#8
Hiroshi Gomi, Hiromi Osawa, Rie Uno, Tadashi Yasui, Masahiro Hosaka, Seiji Torii, Azuma Tsukise
The comparative structure and expression of salivary components and vesicular transport proteins in the canine major salivary glands were investigated. Histochemical analysis revealed that the morphology of the five major salivary glands-parotid, submandibular, polystomatic sublingual, monostomatic sublingual, and zygomatic glands-was greatly diverse. Immunoblot analysis revealed that expression levels of α-amylase and antimicrobial proteins, such as lysozyme, lactoperoxidase, and lactoferrin, differed among the different glands...
September 1, 2017: Journal of Histochemistry and Cytochemistry: Official Journal of the Histochemistry Society
https://www.readbyqxmd.com/read/28900128/the-l-type-voltage-gated-calcium-channel-co-localizes-with-syntaxin-1a-in-nano-clusters-at-the-plasma-membrane
#9
Julia Sajman, Michael Trus, Daphne Atlas, Eilon Sherman
The secretory signal elicited by membrane depolarization traverses from the Ca(2+)-bound α11.2 pore-forming subunit of the L-type Ca(2+)-channel (Cav1.2) to syntaxin 1 A (Sx1A) via an intra-membrane signaling mechanism. Here, we report the use of two-color Photo-Activated-Localization-Microscopy (PALM) to determine the relation between Cav1.2 and Sx1A in single-molecule detail. We observed nanoscale co-clusters of PAmCherry-tagged Sx1A and Dronpa-tagged α11.2 at a ~1:1 ratio. PAmCherry-tagged Sx1A(C145A), or PAmCherry-tagged Sx2, an inactive Cav1...
September 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28887184/chronic-fluoxetine-administration-enhances-synaptic-plasticity-and-increases-functional-dynamics-in-hippocampal-ca3-ca1-synapses
#10
Dina Popova, Eero Castrén, Tomi Taira
Recent studies demonstrate that chronic administration of the widely used antidepressant fluoxetine (FLX) promotes neurogenesis, synaptogenesis and synaptic plasticity in the adult hippocampus, cortex and amygdala. However, the mechanisms underlying these effects and how are they related to the clinical antidepressant efficacy are still poorly understood. We show here that chronic FLX administration decreases hippocampus-associated neophobia in naïve mice. In parallel, electrophysiological recordings in hippocampal CA3-CA1 circuitry revealed that the FLX treatment resulted in increased short- and long-term plasticity likely attributed to changes in presynaptic function...
September 6, 2017: Neuropharmacology
https://www.readbyqxmd.com/read/28880148/reconciling-isothermal-titration-calorimetry-analyses-of-interactions-between-complexin-and-truncated-snare-complexes
#11
Eric A Prinslow, Chad A Brautigam, Josep Rizo
Neurotransmitter release depends on the SNARE complex formed by syntaxin-1, synaptobrevin and SNAP-25, as well as on complexins, which bind to the SNARE complex and play active and inhibitory roles. A crystal structure of a Complexin-I fragment bearing a so-called 'superclamp' mutation bound to a truncated SNARE complex lacking the C-terminus of the synaptobrevin SNARE motif (SNAREΔ60) suggested that an 'accessory' α-helix of Complexin-I inhibits release by inserting into the C-terminus of the SNARE complex...
September 7, 2017: ELife
https://www.readbyqxmd.com/read/28873199/cloning-and-functional-characterization-of-the-hbsyr1-gene-encoding-a-syntaxin-related-protein-in-tibetan-hulless-barley-hordeum-vulgare-l-var-nudum-hk-f
#12
Q J Xu, Y L Wang, Z X Wei, H J Yuan, X Q Zeng, N Tashi
Tibetan barley is a staple food for the natives of Qinghai-Tibet Plateau. Drought causes a reduction in barley production. In this study, the full-length cDNA of a gene encoding a syntaxin-associated protein was cloned from the leaves of a drought-resistant variety of barley, "Himalaya 10"; its expression was evaluated during drought stress and rehydration via real-time PCR. The cloned HbSYR1 cDNA sequence was 1300 bp in length, and included an 840-bp open reading frame that encoded 279 amino acids. Sequence analysis predicted the molecular weight of the encoded protein to be 42...
August 31, 2017: Genetics and Molecular Research: GMR
https://www.readbyqxmd.com/read/28867551/stable-positioning-of-unc13-restricts-synaptic-vesicle-fusion-to-defined-release-sites-to-promote-synchronous-neurotransmission
#13
Suneel Reddy-Alla, Mathias A Böhme, Eric Reynolds, Christina Beis, Andreas T Grasskamp, Malou M Mampell, Marta Maglione, Meida Jusyte, Ulises Rey, Husam Babikir, Anthony W McCarthy, Christine Quentin, Tanja Matkovic, Dominique Dufour Bergeron, Zeeshan Mushtaq, Fabian Göttfert, David Owald, Thorsten Mielke, Stefan W Hell, Stephan J Sigrist, Alexander M Walter
Neural information processing depends on precisely timed, Ca(2+)-activated synaptic vesicle exocytosis from release sites within active zones (AZs), but molecular details are unknown. Here, we identify that the (M)Unc13-family member Unc13A generates release sites and show the physiological relevance of their restrictive AZ targeting. Super-resolution and intravital imaging of Drosophila neuromuscular junctions revealed that (unlike the other release factors Unc18 and Syntaxin-1A) Unc13A was stably and precisely positioned at AZs...
September 13, 2017: Neuron
https://www.readbyqxmd.com/read/28860966/a-stimulation-function-of-synaptotagmin-1-in-ternary-snare-complex-formation-dependent-on-munc18-and-munc13
#14
Yun Li, Shen Wang, Tianzhi Li, Le Zhu, Yuanyuan Xu, Cong Ma
The Ca(2+) sensor synaptotagmin-1 (Syt1) plays an essential function in synaptic exocytosis. Recently, Syt1 has been implicated in synaptic vesicle priming, a maturation step prior to Ca(2+)-triggered membrane fusion that is believed to involve formation of the ternary SNARE complex and require priming proteins Munc18-1 and Munc13-1. However, the mechanisms of Syt1 in synaptic vesicle priming are still unclear. In this study, we found that Syt1 stimulates the transition from the Munc18-1/syntaxin-1 complex to the ternary SNARE complex catalyzed by Munc13-1...
2017: Frontiers in Molecular Neuroscience
https://www.readbyqxmd.com/read/28858288/simultaneous-lipid-and-content-mixing-assays-for-in-vitro-reconstitution-studies-of-synaptic-vesicle-fusion
#15
Xiaoxia Liu, Alpay Burak Seven, Junjie Xu, Victoria Esser, Lijing Su, Cong Ma, Josep Rizo
This protocol describes reconstitution assays to study how the neurotransmitter release machinery triggers Ca(2+)-dependent synaptic vesicle fusion. The assays monitor fusion between proteoliposomes containing the synaptic vesicle SNARE synaptobrevin (with or without the Ca(2+) sensor synaptotagmin-1) and proteoliposomes initially containing the plasma membrane SNAREs syntaxin-1 and soluble NSF attachment protein (SNAP)-25. Lipid mixing (from fluorescence de-quenching of Marina-Blue-labeled lipids) and content mixing (from development of fluorescence resonance energy transfer (FRET) between phycoerythrin-biotin (PhycoE-Biotin) and Cy5-streptavidin trapped in the two proteoliposome populations) are measured simultaneously to ensure that true, nonleaky membrane fusion is monitored...
September 2017: Nature Protocols
https://www.readbyqxmd.com/read/28855648/the-two-pore-channel-tpc1-is-required-for-efficient-protein-processing-through-early-and-recycling-endosomes
#16
Jan Castonguay, Joachim H C Orth, Thomas Müller, Faten Sleman, Christian Grimm, Christian Wahl-Schott, Martin Biel, Robert Theodor Mallmann, Wolfgang Bildl, Uwe Schulte, Norbert Klugbauer
Two-pore channels (TPCs) are localized in endo-lysosomal compartments and assumed to play an important role for vesicular fusion and endosomal trafficking. Recently, it has been shown that both TPC1 and 2 were required for host cell entry and pathogenicity of Ebola viruses. Here, we investigate the cellular function of TPC1 using protein toxins as model substrates for distinct endosomal processing routes. Toxin uptake and activation through early endosomes but not processing through other compartments were reduced in TPC1 knockout cells...
August 30, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28827281/munc18a-clusters-snare-bearing-liposomes-prior-to-trans-snare-zippering
#17
Matthew Grant Arnold, Pratikshya Adhikari, Baobin Kang, Hao Xu 徐昊
Sec1-Munc18 (SM) proteins co-operate with SNAREs {SNAP [soluble NSF (N-ethylmaleimide-sensitive factor) attachment protein] receptors} to mediate membrane fusion in eukaryotic cells. Studies of Munc18a/Munc18-1/Stxbp1 in neurotransmission suggest that SM proteins accelerate fusion kinetics primarily by activating the partially zippered trans-SNARE complex. However, accumulating evidence has argued for additional roles for SM proteins in earlier steps in the fusion cascade. Here, we investigate the function of Munc18a in reconstituted exocytic reactions mediated by neuronal and non-neuronal SNAREs...
September 24, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28815213/identification-of-rab18-as-an-essential-host-factor-for-bk-polyomavirus-infection-using-a-whole-genome-rna-interference-screen
#18
Linbo Zhao, Michael J Imperiale
BK polyomavirus (BKPyV) is a human pathogen first isolated in 1971. BKPyV infection is ubiquitous in the human population, with over 80% of adults worldwide being seropositive for BKPyV. BKPyV infection is usually asymptomatic; however, BKPyV reactivation in immunosuppressed transplant patients causes two diseases, polyomavirus-associated nephropathy and hemorrhagic cystitis. To establish a successful infection in host cells, BKPyV must travel in retrograde transport vesicles to reach the nucleus. To make this happen, BKPyV requires the cooperation of host cell proteins...
July 2017: MSphere
https://www.readbyqxmd.com/read/28814500/monoubiquitination-of-syntaxin-3-leads-to-retrieval-from-the-basolateral-plasma-membrane-and-facilitates-cargo-recruitment-to-exosomes
#19
Adrian J Giovannone, Elena Reales, Pallavi Bhattaram, Alberto Fraile-Ramos, Thomas Weimbs
Syntaxin 3 (Stx3), a SNARE protein located and functioning at the apical plasma membrane of epithelial cells, is required for epithelial polarity. A fraction of Stx3 is localized to late endosomes/lysosomes, although how it traffics there and its function in these organelles is unknown. Here we report that Stx3 undergoes monoubiquitination in a conserved polybasic domain. Stx3 present at the basolateral-but not the apical-plasma membrane is rapidly endocytosed, targeted to endosomes, internalized into intraluminal vesicles (ILVs), and excreted in exosomes...
October 15, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28800600/botulinum-neurotoxin-c-mutants-reveal-different-effects-of-syntaxin-or-snap-25-proteolysis-on-neuromuscular-transmission
#20
Giulia Zanetti, Stefan Sikorra, Andreas Rummel, Nadja Krez, Elisa Duregotti, Samuele Negro, Tina Henke, Ornella Rossetto, Thomas Binz, Marco Pirazzini
Botulinum neurotoxin serotype C (BoNT/C) is a neuroparalytic toxin associated with outbreaks of animal botulism, particularly in birds, and is the only BoNT known to cleave two different SNARE proteins, SNAP-25 and syntaxin. BoNT/C was shown to be a good substitute for BoNT/A1 in human dystonia therapy because of its long lasting effects and absence of neuromuscular damage. Two triple mutants of BoNT/C, namely BoNT/C S51T/R52N/N53P (BoNT/C α-51) and BoNT/C L200W/M221W/I226W (BoNT/C α-3W), were recently reported to selectively cleave syntaxin and have been used here to evaluate the individual contribution of SNAP-25 and syntaxin cleavage to the effect of BoNT/C in vivo...
August 2017: PLoS Pathogens
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