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https://www.readbyqxmd.com/read/28439275/arabidopsis-ala1-and-ala2-mediate-rnai-based-antiviral-immunity
#1
Biyun Zhu, Hua Gao, Gang Xu, Dewei Wu, Susheng Song, Hongshan Jiang, Shuifang Zhu, Tiancong Qi, Daoxin Xie
RNA intereferencing (RNAi) pathway regulates antiviral immunity and mediates plant growth and development. Despite considerable research efforts, a few components in RNAi pathway have been revealed, including ARGONAUTEs (AGOs), DICER-LIKEs (DCLs), RNA-dependent RNA polymerase 1 and 6 (RDR1/6), and ALTERED MERISTEM PROGRAM 1 (AMP1). In this study, we performed a forward genetic screening for enhancers of rdr6 via inoculation of CMV2aTΔ2b, a 2b-deficient Cucumber Mosaic Virus that is unable to suppress RNAi-mediated antiviral immunity...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28434950/an-arabidopsis-lipid-flippase-is-required-for-timely-recruitment-of-defenses-to-the-host-pathogen-interface-at-the-plant-cell-surface
#2
William Underwood, Andrew Ryan, Shauna C Somerville
Deposition of cell wall-reinforcing papillae is an integral component of the plant immune response. The Arabidopsis PENETRATION 3 (PEN3) ATP binding cassette (ABC) transporter plays a role in defense against numerous pathogens and is recruited to sites of pathogen detection where it accumulates within papillae. However, the trafficking pathways and regulatory mechanisms contributing to recruitment of PEN3 and other defenses to the host-pathogen interface are poorly understood. Here, we report a confocal microscopy-based screen to identify mutants with altered localization of PEN3-GFP after inoculation with powdery mildew fungi...
April 18, 2017: Molecular Plant
https://www.readbyqxmd.com/read/28432259/probing-native-metal-ion-association-sites-through-quenching-of-fluorophores-in-the-nucleotide-binding-domains-of-the-abc-transporter-msba
#3
Daiki Tatsumi, Kei Nanatani, Yuto Koike, Kiyoto Kamagata, Satoshi Takahashi, Ayumu Konno, Tadaomi Furuta, Minoru Sakurai, Nobuyuki Uozumi
ATP binding cassette (ABC) transporters are ubiquitously present in prokaryotic and eukaryotic cells. Binding of ATP to the nucleotide binding domains (NBDs) elicits major conformational changes of the transporters resulting in the transport of the substrate across the membrane. The availability of a crystal structure of the NBDs enabled us to elucidate the local structure and small-scale dynamics in the NBDs. Here, we labeled the ABC transporter MsbA, a homodimeric flippase from E. coli , with a fluorescent probe, Alexa532, within the NBDs...
April 21, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28422165/structural-basis-of-inhibition-of-lipid-linked-oligosaccharide-flippase-pglk-by-a-conformational-nanobody
#4
Camilo Perez, Martin Köhler, Daniel Janser, Els Pardon, Jan Steyaert, Renato Zenobi, Kaspar P Locher
PglK is an ABC transporter that flips a lipid-linked oligosaccharide (LLO) that serves as a donor in protein N-glycosylation. Previous structures revealed two inward-facing conformations, both with very large separations of the nucleotide binding domains (NBDs), and a closed, ADP-bound state that featured an occluded cavity. To investigate additional states, we developed conformation-sensitive, single-domain camelid nanobodies (Nb) and studied their effect on PglK activity. Biochemical, structural, and mass spectrometric analyses revealed that one inhibitory Nb binds as a single copy to homodimeric PglK...
April 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28404745/quantitative-high-content-imaging-identifies-novel-regulators-of-neo1-trafficking-at-endosomes
#5
Lauren E Dalton, Björn D M Bean, Michael Davey, Elizabeth Conibear
P4-ATPases are a family of putative phospholipid flippases that regulate lipid membrane asymmetry, which is important for vesicle formation. Two yeast flippases, Drs2 and Neo1, have non-redundant functions in the recycling of the synaptobrevin-like v-SNARE, Snc1, from early endosomes. Drs2 activity is needed to form vesicles and to regulate its own trafficking, suggesting that flippase activity and localization are linked. However, the role of Neo1 in endosomal recycling is not well characterized. To identify novel regulators of Neo1 trafficking and activity at endosomes, we first identified mutants with impaired recycling of a Snc1-based reporter and subsequently used high-content microscopy to classify these mutants based on the localization of Neo1 or its binding partners Mon2 and Dop1...
April 12, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28357373/cryptococcus-flips-its-lid-membrane-phospholipid-asymmetry-modulates-antifungal-drug-resistance-and-virulence
#6
COMMENT
Erika Shor, Yina Wang, David S Perlin, Chaoyang Xue
Human fungal infections are increasing in prevalence and acquisition of antifungal drug resistance, while our antifungal drug armamentarium remains very limited, constituting a significant public health problem. Despite the fact that prominent antifungal drugs target the fungal cell membrane, very little is known about how fungal membrane biology regulates drug-target interactions. Asymmetrical phospholipid distribution is an essential property of biological membranes, which is maintained by a group of transporters that dynamically translocate specific phospholipid groups across the membrane bilayer...
August 1, 2016: Microbial Cell
https://www.readbyqxmd.com/read/28324143/rapid-customised-operon-assembly-by-yeast-recombinational-cloning
#7
Michael A Liu, Johanna J Kenyon, Jason Lee, Peter R Reeves
We have developed a system called the Operon Assembly Protocol (OAP), which takes advantage of the homologous recombination DNA repair pathway in Saccharomyces cerevisiae to assemble full-length operons from a series of overlapping PCR products into a specially engineered yeast-Escherichia coli shuttle vector. This flexible, streamlined system can be used to assemble several operon clones simultaneously, and each clone can be expressed in the same E. coli tester strain to facilitate direct functional comparisons...
March 21, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28302728/a-high-and-phosphatidylinositol-4-phosphate-pi4p-dependent-atpase-activity-for-the-drs2p-cdc50p-flippase-after-removal-of-its-n-and-c-terminal-extensions
#8
Hassina Azouaoui, Cédric Montigny, Thibaud Dieudonné, Philippe Champeil, Aurore Jacquot, José Luis Vázquez-Ibar, Pierre Le Maréchal, Jakob Ulstrup, Miriam-Rose Ash, Joseph A Lyons, Poul Nissen, Guillaume Lenoir
P4-ATPases, also known as phospholipid flippases, are responsible for creating and maintaining transbilayer lipid asymmetry in eukaryotic cell membranes. Here, we use limited proteolysis to investigate the role of the N- and C-termini in ATP hydrolysis and auto-inhibition of the yeast flippase Drs2p/Cdc50p. We show that limited proteolysis of the detergent-solubilized and purified yeast flippase may result in more than one order of magnitude increase of its ATPase activity, which remains dependent on phosphatidylinositol-4-phosphate (PI4P), a regulator of this lipid flippase, and specific to a phosphatidylserine substrate...
March 16, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28288918/increased-carbon-dioxide-levels-stimulate-neutrophils-to-produce-microparticles-and-activate-the-nucleotide-binding-domain-like-receptor-3-inflammasome
#9
Stephen R Thom, Veena M Bhopale, JingPing Hu, Ming Yang
We hypothesized that elevations of carbon dioxide (CO2) commonly found in modern buildings will stimulate leukocytes to produce microparticles (MPs) and activate the nucleotide-binding domain-like receptor 3 (NLRP3) inflammasome due to mitochondrial oxidative stress. Human and murine neutrophils generate MPs with high interleukin-1β (IL-1β) content when incubated ex vivo in buffer equilibrated with 0.1-0.4% additional CO2. Enhanced MPs production requires mitochondrial reactive oxygen species production, which is mediated by activities of pyruvate carboxylase and phosphoenolpyruvate carboxykinase...
March 10, 2017: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/28272479/plasmalogen-biosynthesis-is-spatiotemporally-regulated-by-sensing-plasmalogens-in-the-inner-leaflet-of-plasma-membranes
#10
Masanori Honsho, Yuichi Abe, Yukio Fujiki
Alkenyl ether phospholipids are a major sub-class of ethanolamine- and choline-phospholipids in which a long chain fatty alcohol is attached at the sn-1 position through a vinyl ether bond. Biosynthesis of ethanolamine-containing alkenyl ether phospholipids, plasmalogens, is regulated by modulating the stability of fatty acyl-CoA reductase 1 (Far1) in a manner dependent on the level of cellular plasmalogens. However, precise molecular mechanisms underlying the regulation of plasmalogen synthesis remain poorly understood...
March 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28253836/flippant-an-r-package-for-the-automated-analysis-of-fluorescence-based-scramblase-assays
#11
Richard J Cotton, Birgit Ploier, Michael A Goren, Anant K Menon, Johannes Graumann
BACKGROUND: The lipid scrambling activity of protein extracts and purified scramblases is typically measured using a fluorescence-based assay. While the assay has yielded insight into the scramblase activity in crude membrane preparations, functional validation of candidate scramblases, stoichiometry of scramblase complexes as well as ATP-dependence of flippases, data analysis in its context has remained a task involving many manual steps. RESULTS: With the extension package "flippant" to R, a free software environment for statistical computing and graphics, we introduce an integrated solution for the analysis and publication-grade graphical presentation of dithionite scramblase assays and demonstrate its utility in revisiting an originally manual analysis from the publication record, closely reproducing the reported results...
March 3, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28220754/the-ap-2-complex-has-a-specialized-clathrin-independent-role-in-apical-endocytosis-and-polar-growth-in-fungi
#12
Olga Martzoukou, Sotiris Amillis, Amalia Zervakou, Savvas Christoforidis, George Diallinas
Filamentous fungi provide excellent systems for investigating the role of the AP-2 complex in polar growth. Using Aspergillus nidulans, we show that AP-2 has a clathrin-independent essential role in polarity maintenance and growth. This is in line with a sequence analysis showing that the AP-2 β subunit (β2) of higher fungi lacks a clathrin-binding domain, and experiments showing that AP-2 does not co-localize with clathrin. We provide genetic and cellular evidence that AP-2 interacts with endocytic markers SlaB(End4) and SagA(End3) and the lipid flippases DnfA and DnfB in the sub-apical collar region of hyphae...
February 21, 2017: ELife
https://www.readbyqxmd.com/read/28175960/erratum-to-phospholipid-flippases-attenuate-lps-induced-tlr4-signaling-by-mediating-endocytic-retrieval-of-toll-like-receptor-4
#13
Vincent A van der Mark, Mohammed Ghiboub, Casper Marsman, Jing Zhao, Remco van Dijk, Johan K Hiralall, Kam S Ho-Mok, Zoë Castricum, Wouter J de Jonge, Ronald P J Oude Elferink, Coen C Paulusma
No abstract text is available yet for this article.
April 2017: Cellular and Molecular Life Sciences: CMLS
https://www.readbyqxmd.com/read/28167678/ypk1-ypk2-kinases-maintain-rho1-at-the-plasma-membrane-by-flippase-dependent-lipid-remodelling-after-membrane-stresses
#14
Riko Hatakeyama, Keiko Kono, Satoshi Yoshida
The plasma membrane (PM) is frequently challenged by mechanical stresses. In budding yeast, TORC2-Ypk1/Ypk2 kinase cascade plays a critical role in PM stress responses by reorganizing the actin cytoskeleton via Rho1 GTPase. However, the molecular mechanism by which TORC2-Ypk1/Ypk2 regulates Rho1 is not well defined. Here, we found that Ypk1/Ypk2 maintain PM localization of Rho1 under PM stress via spatial reorganization of the lipids including phosphatidylserine (PS). Genetic evidence suggests that this process is mediated by the Lem3-containing lipid flippase...
February 6, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28123063/lipid-flippases-promote-antiviral-silencing-and-the-biogenesis-of-viral-and-host-sirnas-in-arabidopsis
#15
Zhongxin Guo, Jinfeng Lu, Xianbing Wang, Binhui Zhan, Wanxiang Li, Shou-Wei Ding
Dicer-mediated processing of virus-specific dsRNA into short interfering RNAs (siRNAs) in plants and animals initiates a specific antiviral defense by RNA interference (RNAi). In this study, we developed a forward genetic screen for the identification of host factors required for antiviral RNAi in Arabidopsis thaliana Using whole-genome sequencing and a computational pipeline, we identified aminophospholipid transporting ATPase 2 (ALA2) and the related ALA1 in the type IV subfamily of P-type ATPases as key components of antiviral RNAi...
February 7, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28118404/slow-phospholipid-exchange-between-a-detergent-solubilized-membrane-protein-and-lipid-detergent-mixed-micelles-brominated-phospholipids-as-tools-to-follow-its-kinetics
#16
Cédric Montigny, Thibaud Dieudonné, Stéphane Orlowski, José Luis Vázquez-Ibar, Carole Gauron, Dominique Georgin, Sten Lund, Marc le Maire, Jesper V Møller, Philippe Champeil, Guillaume Lenoir
Membrane proteins are largely dependent for their function on the phospholipids present in their immediate environment, and when they are solubilized by detergent for further study, residual phospholipids are critical, too. Here, brominated phosphatidylcholine, a phospholipid which behaves as an unsaturated phosphatidylcholine, was used to reveal the kinetics of phospholipid exchange or transfer from detergent mixed micelles to the environment of a detergent-solubilized membrane protein, the paradigmatic P-type ATPase SERCA1a, in which Trp residues can experience fluorescence quenching by bromine atoms present on phospholipid alkyl chains in their immediate environment...
2017: PloS One
https://www.readbyqxmd.com/read/28118222/maintenance-and-regulation-of-asymmetric-phospholipid-distribution-in-human-erythrocyte-membranes-implications-for-erythrocyte-functions
#17
Nobuto Arashiki, Yuichi Takakuwa
PURPOSE OF REVIEW: The article summarizes new insights into the molecular mechanisms for the maintenance and regulation of the asymmetric distribution of phospholipids in human erythrocyte membranes. We focus on phosphatidylserine, which is primarily found in the inner leaflet of the membrane lipid bilayer under low Ca conditions (<1 μmol/l) and is exposed to the outer leaflet under elevated Ca concentrations (>1 μmol/l), when cells become senescent. Clarification of the molecular basis of phosphatidylserine flipping and scrambling is important for addressing long-standing questions regarding phosphatidylserine functions...
May 2017: Current Opinion in Hematology
https://www.readbyqxmd.com/read/28114371/correction-the-o-antigen-flippase-wzk-can-substitute-for-murj-in-peptidoglycan-synthesis-in-helicobacter-pylori-and-escherichia-coli
#18
Wael Elhenawy, Rebecca M Davis, Jutta Fero, Nina R Salama, Mario F Feldman, Natividad Ruiz
[This corrects the article DOI: 10.1371/journal.pone.0161587.].
2017: PloS One
https://www.readbyqxmd.com/read/28109698/microbubble-assisted-ultrasound-induced-transient-phosphatidylserine-translocation
#19
Jean-Michel Escoffre, Marc Derieppe, Bart Lammertink, Clemens Bos, Chrit Moonen
Microbubble-assisted ultrasound (sonopermeabilization) results in reversible permeabilization of the plasma membrane of cells. This method is increasingly used in vivo because of its potential to deliver therapeutic molecules with limited cell damage. Nevertheless, the effects of sonopermeabilization on the plasma membrane remain not fully understood. We investigated the influence of sonopermeabilization on the transverse mobility of phospholipids, especially on phosphatidylserine (PS) externalization. We performed studies using optical imaging with Annexin V and FM1-43 probes to monitor PS externalization of rat glioma C6 cells...
April 2017: Ultrasound in Medicine & Biology
https://www.readbyqxmd.com/read/28069741/tor-complex-2-regulated-protein-kinase-fpk1-stimulates-endocytosis-via-inhibition-of-ark1-prk1-related-protein-kinase-akl1-in-saccharomyces-cerevisiae
#20
Françoise M Roelants, Kristin L Leskoske, Ross T A Pedersen, Alexander Muir, Jeffrey M-H Liu, Gregory C Finnigan, Jeremy Thorner
Depending on the stress, plasma membrane alterations activate or inhibit yeast target of rapamycin (TOR) complex 2, which, in turn, upregulates or downregulates the activity of its essential downstream effector, protein kinase Ypk1. Through phosphorylation of multiple substrates, Ypk1 controls many processes that restore homeostasis. One such substrate is protein kinase Fpk1, which is negatively regulated by Ypk1. Fpk1 phosphorylates and stimulates flippases that translocate aminoglycerophospholipids from the outer to the inner leaflet of the plasma membrane...
April 1, 2017: Molecular and Cellular Biology
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