DOCSLIB.ORG

Small Genome Symbiont Underlies Cuticle Hardness in Beetles

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

Small genome symbiont underlies cuticle hardness PNAS PLUS in beetles Hisashi Anbutsua,b,1,2, Minoru Moriyamaa,1, Naruo Nikohc,1, Takahiro Hosokawaa,d, Ryo Futahashia, Masahiko Tanahashia, Xian-Ying Menga, Takashi Kuriwadae,f, Naoki Morig, Kenshiro Oshimah, Masahira Hattorih,i, Manabu Fujiej, Noriyuki Satohk, Taro Maedal, Shuji Shigenobul, Ryuichi Kogaa, and Takema Fukatsua,m,n,2 aBioproduction Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba 305-8566, Japan; bComputational Bio Big-Data Open Innovation Laboratory, National Institute of Advanced Industrial Science and Technology, Tokyo 169-8555, Japan; cDepartment of Liberal Arts, The Open University of Japan, Chiba 261-8586, Japan; dFaculty of Science, Kyushu University, Fukuoka 819-0395, Japan; eNational Agriculture and Food Research Organization, Kyushu Okinawa Agricultural Research Center, Okinawa 901-0336, Japan; fFaculty of Education, Kagoshima University, Kagoshima 890-0065, Japan; gDivision of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan; hGraduate School of Frontier Sciences, University of Tokyo, Chiba 277-8561, Japan; iGraduate School of Advanced Science and Engineering, Waseda University, Tokyo 169-8555, Japan; jDNA Sequencing Section, Okinawa Institute of Science and Technology Graduate University, Okinawa 904-0495, Japan; kMarine Genomics Unit, Okinawa Institute of Science and Technology Graduate University, Okinawa 904-0495, Japan; lNIBB Core Research Facilities, National Institute for Basic Biology, Okazaki 444-8585, Japan; mDepartment of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan; and nGraduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba 305-8572, Japan Edited by Nancy A. Moran, University of Texas at Austin, Austin, TX, and approved August 28, 2017 (received for review July 19, 2017) Beetles, representing the majority of the insect species diversity, are symbiont transmission over evolutionary time (4, 6, 7). Some bac- characterized by thick and hard cuticle, which plays important roles terial symbionts of plant-sucking insects like cicadas, leafhoppers, for their environmental adaptation and underpins their inordinate spittlebugs, psyllids, and mealybugs belonging to the order Hemi- diversity and prosperity. Here, we report a bacterial endosymbiont ptera, which are associated with multiple endocellular bacterial extremely specialized for sustaining beetle’s cuticle formation. cosymbionts within the symbiotic organ called the bacteriome, have γ Many weevils are associated with a -proteobacterial endosymbiont experienced extreme genome reduction down to 0.2 Mb or smaller Nardonella lineage , whose evolutionary origin is estimated as older with 200 or less protein-coding genes (8–13), suggesting that met- EVOLUTION than 100 million years, but its functional aspect has been elusive. abolic complementation between the cosymbionts may have fur- Sequencing of Nardonella genomes from diverse weevils unveiled ther facilitated the reductive genome evolution entailing losses of drastic size reduction to 0.2 Mb, in which minimal complete gene – sets for bacterial replication, transcription, and translation were pre- otherwise essential genes in either of the cosymbionts (7, 14 17). In this study, we report another case of extremely reduced sent but almost all of the other metabolic pathway genes were missing. Notably, the only metabolic pathway retained in the symbiont genome in a different insect group through a different Nardonella genomes was the tyrosine synthesis pathway, identifying evolutionary trajectory. Beetles, comprising the insect order tyrosine provisioning as Nardonella’s sole biological role. Weevils are Coleoptera, represent the majority of the biodiversity described armored with hard cuticle, tyrosine is the principal precursor for cu- ticle formation, and experimental suppression of Nardonella resulted Significance in emergence of reddish and soft weevils with low tyrosine titer, confirming the importance of Nardonella-mediated tyrosine produc- Beetles are successful in the terrestrial ecosystem, which is tion for host’s cuticle formation and hardening. Notably, Nardonella’s attributable to, at least partly, their highly sclerotized exo- tyrosine synthesis pathway was incomplete, lacking the final step skeleton. Here, we report a bacterial symbiont extremely spe- transaminase gene. RNA sequencing identified host’s aminotransfer- cialized for underpinning the beetle’s hardness. The ancient ase genes up-regulated in the bacteriome. RNA interference target- endosymbiont Nardonella associated with weevils has an ex- ing the aminotransferase genes induced reddish and soft weevils tremely small genome devoted to a single biological function, with low tyrosine titer, verifying host’s final step regulation of the tyrosine provisioning, which is needed for insect’s cuticle for- tyrosine synthesis pathway. Our finding highlights an impressively mation and hardening. Notably, only the final step reaction of intimate and focused aspect of the host–symbiont metabolic integ- the tyrosine synthesis pathway is complemented by host- rity via streamlined evolution for a single biological function of encoded aminotransferases up-regulated in the bacteriome, ecological relevance. highlighting a highly focused aspect of the host–symbiont metabolic integrity. Both symbiont suppression by an antibiotic weevil | Nardonella | symbiont | genome | tyrosine and RNA interference of the host aminotransferases induce reddish and soft weevils, verifying the pivotal role of the ymbiotic associations with microorganisms are ubiquitously symbiosis for the beetle’s hardness. Sfound in a variety of insects, which are rated among the im- portant factors underpinning their adaptation, diversity, and pros- Author contributions: H.A., M.M., N.N., T.H., and T.F. designed research; H.A., N.N., T.H., – M.T., X.-Y.M., T.K., N.M., K.O., M.F., T.M., and R.K. performed research; M.H., N.S., and S.S. perity (1 3). Many bacterial symbionts are indispensable for contributed new reagents/analytic tools; H.A., M.M., N.N., T.H., R.F., M.T., K.O., M.F., and growth, survival, and reproduction of their insect hosts via, for T.M. analyzed data; and H.A., M.M., N.N., R.F., and T.F. wrote the paper. example, provisioning of essential nutrients like amino acids and The authors declare no conflict of interest. vitamins, where the host and the symbiont are integrated into an This article is a PNAS Direct Submission. almost inseparable biological entity (4, 5). In such obligate symbi- Freely available online through the PNAS open access option. otic associations, the symbiont genomes tend to exhibit conspicu- Data deposition: The sequences reported in this paper have been deposited in the DNA ous structural degeneration, massive gene losses, and drastic size Data Bank Japan Read Archive, www.ddbj.nig.ac.jp (accession nos. AP018159–AP018162, reduction, which are attributable to relaxed natural selection acting LC260175–LC260180, LC260491, and DRR095964–DRR095975). on many symbiont genes no longer necessary for the permanent 1H.A., M.M. and N.N. contributed equally to this work. intrahost lifestyle, and also to accumulation of deleterious muta- 2To whom correspondence may be addressed. Email: [email protected] or t-fukatsu@ tions driven by attenuated natural selection acting on the symbiont aist.go.jp. genomes due to strong population bottlenecks and restricted This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. horizontal gene acquisitions associated with continuous vertical 1073/pnas.1712857114/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1712857114 PNAS Early Edition | 1of10 Downloaded by guest on September 26, 2021 – ’ A C lasting host symbiont coevolution, Nardonella s biological role Bacteriome has been poorly understood (26). Previous studies have identi- fied a number of weevil lineages in which Nardonella infections have been lost or replaced by different bacterial lineages, uncovering a strikingly dynamic aspect of the endosymbiotic evolution in the insect group (23, 24, 32–35). Here, we report genomic, transcriptomic, and functional 1 cm analyses of the Nardonella symbionts associated with diverse 5 mm weevils, which unveiled their extremely reduced genomes down B to as small as 0.2 Mb in the absence of any cosymbionts. The tiny Midgut genomes encode minimal but complete gene sets for bacterial 500 m Foregut replication, transcription, and translation, while lacking almost E all of the other metabolic pathway genes, which indicate Nar- donella’s near-complete dependence on host-derived metabolites D toward a minimal cellular entity through the ancient coevolu- tionary history. Notably, a set of metabolic genes is conspicu- ously retained in the Nardonella genomes, namely synthesis pathway genes for a specific amino acid, tyrosine. Weevils are Oocyte armored with hard cuticle, tyrosine is the principal precursor 1 mm needed for cuticle formation, and the Nardonella genome has G 1 m been streamlined for a single biological function, tyrosine pro- 200 µm visioning, for sustaining the weevil’s highly sclerotized exoskeleton, which elucidates the general importance of endosymbiont- F provisioned tyrosine for cuticle formation in weevils (36, 37), and potentially also in other beetles. Furthermore, we demon- strate that, reflecting the absence of transcriptional regulators in the tiny symbiont
Recommended publications
  • Aspartate Aminotransferase (AST, GOT), Human Liver

    Aspartate Aminotransferase (AST, GOT), Human Liver

    BioVision 05/18 For research use only Aspartate Aminotransferase (AST, GOT), Human Liver CATALOG NO: P1299-100 1 units RELATED PRODUCTS: ALTERNATE NAMES: Aspartate Transaminase, Glutamate Oxaloacetate, AST, GOT, Aspartate Aminotransferase (AST) (Mouse) ELISA Kit (Cat. No. E4320) sGOT, AspAT, ASAT, serum glutamic oxaloacetic transaminase, AAT Aspartate Aminotransferase (AST) (Human) ELISA Kit (Cat. No. E4319) Aspartate Aminotransferase (AST) (Rat) ELISA Kit (Cat. No. E4321) SOURCE: Human Liver Aspartate Aminotransferase (AST or SGOT) Activity Colorimetric Assay Kit (Cat. PURITY: Purified No. K753) Anti-GOT2 Antibody (cat. No. A1273) MOL. WEIGHT: ~92 kDa Anti-GOT1 Antibody (Cat. No. A1272) FORM: Lyophilized GOT2, human recombinant (Cat. No. 7809) GOT1, human recombinant (Cat. No. 7808) STORAGE CONDITIONS: Store at -20°C. Avoid repeated freezing and thawing cycles. BIOLOGICAL ACTIVITY: ≥ 1 U/mg (Dimension® Clinical Chemistry System) UNIT DEFINITATION: One unit will catalyze the transamination of one micromole of L- aspartate to alpha-ketoglutarate forming L-glutamate and oxaloacetate per minute at 37°C and pH 7.8.Measured at 340 nm as one equimolar amount of NAD produced by a coupled reaction. RECONSTITUTION: > 1 mg/mL in tris buffered saline, 1% BSA, pH 8.0. AST/SGOT is found in many tissues throughout the body, including DESCRIPTION: the liver, heart, muscles, kidney, and brain. If any of these organs or tissues is affected by disease or injury, AST is released into the bloodstream. This means that AST isn't as specific an indicator of liver damage as ALT (also known as alanine aminotransferase, another type of enzyme found almost entirely in the liver). The ratio of AST and ALT levels are commonly used as biomarkers for liver health.
  • Supplemental Figure 1. Vimentin

    Supplemental Figure 1. Vimentin

    Double mutant specific genes Transcript gene_assignment Gene Symbol RefSeq FDR Fold- FDR Fold- FDR Fold- ID (single vs. Change (double Change (double Change wt) (single vs. wt) (double vs. single) (double vs. wt) vs. wt) vs. single) 10485013 BC085239 // 1110051M20Rik // RIKEN cDNA 1110051M20 gene // 2 E1 // 228356 /// NM 1110051M20Ri BC085239 0.164013 -1.38517 0.0345128 -2.24228 0.154535 -1.61877 k 10358717 NM_197990 // 1700025G04Rik // RIKEN cDNA 1700025G04 gene // 1 G2 // 69399 /// BC 1700025G04Rik NM_197990 0.142593 -1.37878 0.0212926 -3.13385 0.093068 -2.27291 10358713 NM_197990 // 1700025G04Rik // RIKEN cDNA 1700025G04 gene // 1 G2 // 69399 1700025G04Rik NM_197990 0.0655213 -1.71563 0.0222468 -2.32498 0.166843 -1.35517 10481312 NM_027283 // 1700026L06Rik // RIKEN cDNA 1700026L06 gene // 2 A3 // 69987 /// EN 1700026L06Rik NM_027283 0.0503754 -1.46385 0.0140999 -2.19537 0.0825609 -1.49972 10351465 BC150846 // 1700084C01Rik // RIKEN cDNA 1700084C01 gene // 1 H3 // 78465 /// NM_ 1700084C01Rik BC150846 0.107391 -1.5916 0.0385418 -2.05801 0.295457 -1.29305 10569654 AK007416 // 1810010D01Rik // RIKEN cDNA 1810010D01 gene // 7 F5 // 381935 /// XR 1810010D01Rik AK007416 0.145576 1.69432 0.0476957 2.51662 0.288571 1.48533 10508883 NM_001083916 // 1810019J16Rik // RIKEN cDNA 1810019J16 gene // 4 D2.3 // 69073 / 1810019J16Rik NM_001083916 0.0533206 1.57139 0.0145433 2.56417 0.0836674 1.63179 10585282 ENSMUST00000050829 // 2010007H06Rik // RIKEN cDNA 2010007H06 gene // --- // 6984 2010007H06Rik ENSMUST00000050829 0.129914 -1.71998 0.0434862 -2.51672
  • Ornithine Aminotransferase, an Important Glutamate-Metabolizing Enzyme at the Crossroads of Multiple Metabolic Pathways

    Ornithine Aminotransferase, an Important Glutamate-Metabolizing Enzyme at the Crossroads of Multiple Metabolic Pathways

    biology Review Ornithine Aminotransferase, an Important Glutamate-Metabolizing Enzyme at the Crossroads of Multiple Metabolic Pathways Antonin Ginguay 1,2, Luc Cynober 1,2,*, Emmanuel Curis 3,4,5,6 and Ioannis Nicolis 3,7 1 Clinical Chemistry, Cochin Hospital, GH HUPC, AP-HP, 75014 Paris, France; [email protected] 2 Laboratory of Biological Nutrition, EA 4466 PRETRAM, Faculté de Pharmacie, Université Paris Descartes, 75006 Paris, France 3 Laboratoire de biomathématiques, plateau iB2, Faculté de Pharmacie, Université Paris Descartes, 75006 Paris, France; [email protected] (E.C.); [email protected] (I.N.) 4 UMR 1144, INSERM, Université Paris Descartes, 75006 Paris, France 5 UMR 1144, Université Paris Descartes, 75006 Paris, France 6 Service de biostatistiques et d’informatique médicales, hôpital Saint-Louis, Assistance publique-hôpitaux de Paris, 75010 Paris, France 7 EA 4064 “Épidémiologie environnementale: Impact sanitaire des pollutions”, Faculté de Pharmacie, Université Paris Descartes, 75006 Paris, France * Correspondence: [email protected]; Tel.: +33-158-411-599 Academic Editors: Arthur J.L. Cooper and Thomas M. Jeitner Received: 26 October 2016; Accepted: 24 February 2017; Published: 6 March 2017 Abstract: Ornithine δ-aminotransferase (OAT, E.C. 2.6.1.13) catalyzes the transfer of the δ-amino group from ornithine (Orn) to α-ketoglutarate (aKG), yielding glutamate-5-semialdehyde and glutamate (Glu), and vice versa. In mammals, OAT is a mitochondrial enzyme, mainly located in the liver, intestine, brain, and kidney. In general, OAT serves to form glutamate from ornithine, with the notable exception of the intestine, where citrulline (Cit) or arginine (Arg) are end products.
  • Multiomics Integration Elucidates Metabolic Modulators of Drug

    Multiomics Integration Elucidates Metabolic Modulators of Drug

    bioRxiv preprint doi: https://doi.org/10.1101/2021.01.07.425721; this version posted January 8, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Multiomics Integration Elucidates Metabolic Modulators of Drug Resistance in Lymphoma Choueiry, Fouad1*, Singh, Satishkumar2,3*, Sun, Xiaowei1, Zhang, Shiqi1, Sircar, Anuvrat2,3 ,Hart, Amber2, Alinari, Lapo2,3, Narendranath Epperla2,3, Baiocchi, Robert2,3, Zhu, Jiangjiang1,# and Sehgal, Lalit2,3# 1 Department of Human Sciences, The Ohio State University, Columbus, OH 43210; 2 Division of Hematology, Department of Internal Medicine, The Ohio State, 3 James Comprehensive Cancer Center, The Ohio State University Columbus, OH 43210. *Equal contribution, #Corresponding authors Abstract Background Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma (NHL). B-cell NHLs rely on Bruton’s tyrosine kinase (BTK) mediated B-cell receptor signaling for survival and disease progression. However, they are often resistant to BTK inhibitors or soon acquire resistance after drug exposure resulting in the drug tolerant form. The drug tolerant clones proliferate faster, have increased metabolic activity, and shift to oxidative phosphorylation; however, how this metabolic programming occurs in the drug resistant tumor is poorly understood. Methods In this study, we explored for the first time the metabolic regulators of ibrutinib-resistant activated B-cell (ABC) DLBCL using a ‘multi-omics’ analysis that integrated metabolomics (using high-resolution mass spectrometry) and transcriptomic (gene expression analysis). Overlay of the unbiased statistical analyses, genetic perturbation and pharmaceutical inhibition, were further used to identify the key players that contribute to the metabolic reprograming of the drug resistant clone.
  • Insect Imaging at the ANKA Synchrotron Radiation Facility 147

    Insect Imaging at the ANKA Synchrotron Radiation Facility 147

    Insect Imaging at the ANKA Synchrotron Radiation Facility 147 Entomologie heute 25 (2013): 147-160 Insect Imaging at the ANKA Synchrotron Radiation Facility Bildgebung von Insekten an der Synchrotronstrahlungsquelle ANKA THOMAS VAN DE KAMP, ALEXEY ERSHOV, TOMY DOS SANTOS ROLO, ALEXANDER RIEDEL & TILO BAUMBACH Summary: Internal structures of biological samples are often diffi cult to visualize by traditional morphological methods like light and electron microscopy. In insects, a robust cuticle often impedes examination. Three-dimensional visualization of anatomical details based on light microscopy photo graphs is particularly challenging, because the necessary creation of a series of “perfect” slices often proves to be impossible in the case of hard-shelled specimens. Synchrotron-based X-ray imaging provides a pool of techniques well-suited for morphological studies. As it allows examin- ing millimeter-sized non-translucent objects, it is particularly valuable for the multidimensional visualization of insects and became increasingly popular among entomologists. A synchrotron is a cyclic particle accelerator. In high vacuum electrons are accelerated up to nearly light speed, injected into a storage ring and deviated by bending magnets. When the electron beam changes its direction due to magnetic infl uence, electromagnetic radiation is transmitted tangentially, which is used in attached experimental stations (“beamlines”). Synchrotron radiation has a broad spectrum ranging from microwaves to hard X-rays, the latter being used for most synchrotron imaging techniques. The high intensity of these X-rays facilitates high special and temporal resolutions. An important method is synchrotron X-ray microtomography (SR-μCT). Here, a series of 2D X-ray projections of a rotating sample is used to calculate a 3D volume.
  • Supplementary Materials

    Supplementary Materials

    1 Supplementary Materials: Supplemental Figure 1. Gene expression profiles of kidneys in the Fcgr2b-/- and Fcgr2b-/-. Stinggt/gt mice. (A) A heat map of microarray data show the genes that significantly changed up to 2 fold compared between Fcgr2b-/- and Fcgr2b-/-. Stinggt/gt mice (N=4 mice per group; p<0.05). Data show in log2 (sample/wild-type). 2 Supplemental Figure 2. Sting signaling is essential for immuno-phenotypes of the Fcgr2b-/-lupus mice. (A-C) Flow cytometry analysis of splenocytes isolated from wild-type, Fcgr2b-/- and Fcgr2b-/-. Stinggt/gt mice at the age of 6-7 months (N= 13-14 per group). Data shown in the percentage of (A) CD4+ ICOS+ cells, (B) B220+ I-Ab+ cells and (C) CD138+ cells. Data show as mean ± SEM (*p < 0.05, **p<0.01 and ***p<0.001). 3 Supplemental Figure 3. Phenotypes of Sting activated dendritic cells. (A) Representative of western blot analysis from immunoprecipitation with Sting of Fcgr2b-/- mice (N= 4). The band was shown in STING protein of activated BMDC with DMXAA at 0, 3 and 6 hr. and phosphorylation of STING at Ser357. (B) Mass spectra of phosphorylation of STING at Ser357 of activated BMDC from Fcgr2b-/- mice after stimulated with DMXAA for 3 hour and followed by immunoprecipitation with STING. (C) Sting-activated BMDC were co-cultured with LYN inhibitor PP2 and analyzed by flow cytometry, which showed the mean fluorescence intensity (MFI) of IAb expressing DC (N = 3 mice per group). 4 Supplemental Table 1. Lists of up and down of regulated proteins Accession No.
  • Supporting Information

    Supporting Information

    Supporting Information Edgar et al. 10.1073/pnas.1601895113 SI Methods (Actimetrics), and recordings were analyzed using LumiCycle Mice. Sample size was determined using the resource equation: Data Analysis software (Actimetrics). E (degrees of freedom in ANOVA) = (total number of exper- – Cell Cycle Analysis of Confluent Cell Monolayers. NIH 3T3, primary imental animals) (number of experimental groups), with −/− sample size adhering to the condition 10 < E < 20. For com- WT, and Bmal1 fibroblasts were sequentially transduced − − parison of MuHV-4 and HSV-1 infection in WT vs. Bmal1 / with lentiviral fluorescent ubiquitin-based cell cycle indicators mice at ZT7 (Fig. 2), the investigator did not know the genotype (FUCCI) mCherry::Cdt1 and amCyan::Geminin reporters (32). of the animals when conducting infections, bioluminescence Dual reporter-positive cells were selected by FACS (Influx Cell imaging, and quantification. For bioluminescence imaging, Sorter; BD Biosciences) and seeded onto 35-mm dishes for mice were injected intraperitoneally with endotoxin-free lucif- subsequent analysis. To confirm that expression of mCherry:: Cdt1 and amCyan::Geminin correspond to G1 (2n DNA con- erin (Promega E6552) using 2 mg total per mouse. Following < ≤ anesthesia with isofluorane, they were scanned with an IVIS tent) and S/G2 (2 n 4 DNA content) cell cycle phases, Lumina (Caliper Life Sciences), 15 min after luciferin admin- respectively, cells were stained with DNA dye DRAQ5 (abcam) and analyzed by flow cytometry (LSR-Fortessa; BD Biosci- istration. Signal intensity was quantified using Living Image ences). To examine dynamics of replicative activity under ex- software (Caliper Life Sciences), obtaining maximum radiance perimental confluent conditions, synchronized FUCCI reporter for designated regions of interest (photons per second per − − − monolayers were observed by time-lapse live cell imaging over square centimeter per Steradian: photons·s 1·cm 2·sr 1), relative 3 d (Nikon Eclipse Ti-E inverted epifluorescent microscope).
  • IL21R Expressing CD14+CD16+ Monocytes Expand in Multiple

    IL21R Expressing CD14+CD16+ Monocytes Expand in Multiple

    Plasma Cell Disorders SUPPLEMENTARY APPENDIX IL21R expressing CD14 +CD16 + monocytes expand in multiple myeloma patients leading to increased osteoclasts Marina Bolzoni, 1 Domenica Ronchetti, 2,3 Paola Storti, 1,4 Gaetano Donofrio, 5 Valentina Marchica, 1,4 Federica Costa, 1 Luca Agnelli, 2,3 Denise Toscani, 1 Rosanna Vescovini, 1 Katia Todoerti, 6 Sabrina Bonomini, 7 Gabriella Sammarelli, 1,7 Andrea Vecchi, 8 Daniela Guasco, 1 Fabrizio Accardi, 1,7 Benedetta Dalla Palma, 1,7 Barbara Gamberi, 9 Carlo Ferrari, 8 Antonino Neri, 2,3 Franco Aversa 1,4,7 and Nicola Giuliani 1,4,7 1Myeloma Unit, Dept. of Medicine and Surgery, University of Parma; 2Dept. of Oncology and Hemato-Oncology, University of Milan; 3Hematology Unit, “Fondazione IRCCS Ca’ Granda”, Ospedale Maggiore Policlinico, Milan; 4CoreLab, University Hospital of Parma; 5Dept. of Medical-Veterinary Science, University of Parma; 6Laboratory of Pre-clinical and Translational Research, IRCCS-CROB, Referral Cancer Center of Basilicata, Rionero in Vulture; 7Hematology and BMT Center, University Hospital of Parma; 8Infectious Disease Unit, University Hospital of Parma and 9“Dip. Oncologico e Tecnologie Avanzate”, IRCCS Arcispedale Santa Maria Nuova, Reggio Emilia, Italy ©2017 Ferrata Storti Foundation. This is an open-access paper. doi:10.3324/haematol. 2016.153841 Received: August 5, 2016. Accepted: December 23, 2016. Pre-published: January 5, 2017. Correspondence: [email protected] SUPPLEMENTAL METHODS Immunophenotype of BM CD14+ in patients with monoclonal gammopathies. Briefly, 100 μl of total BM aspirate was incubated in the dark with anti-human HLA-DR-PE (clone L243; BD), anti-human CD14-PerCP-Cy 5.5, anti-human CD16-PE-Cy7 (clone B73.1; BD) and anti-human CD45-APC-H 7 (clone 2D1; BD) for 20 min.
  • Metabolomics Analysis Reveals Differential T-Cell Serine Metabolism As a Target in Autoimmunity

    Metabolomics Analysis Reveals Differential T-Cell Serine Metabolism As a Target in Autoimmunity

    Metabolomics Analysis Reveals Differential T-Cell Serine Metabolism as a Target in Autoimmunity Gabriela Andrejeva Jeffrey Rathmell lab Vanderbilt University Medical Center [email protected] 2018 UK Metabolism Symposium Monday, 30th July 2018 Outline 2 dUMP dTMP Betaine aldehyde Choline TYMS CHDH Folate Se-Adenosylselenomethionine S-Adenosyl-L-methionine Dihydropteroate Dihydrofolate Cob(II)alamin 10-Formyltetrahydrofolylpolyglutamate10-Formyltetrahydrofolyl Betaine L-glutamate MAT2B MAT1A DHFR MAT2A Tetrahydrofolyl-[Glu](n) Formate Poly-L-glutamate ALDH1L2 MTR 3-Phospho-D-glycerate BHMT2 ALDH1L1 BHMT FPGS S-Adenosyl-L-homocysteine MTHFD1L GGH PHGDH MTHFD1 ATIC AHCYL1 L-Methionine MTHFD2 10-Formyltetrahydrofolate L-Homocysteine MTFMT "N,N-Dimethylglycine" Tetrahydrofolate 3-Phosphonooxypyruvate PDPR AHCY 5-Methyltetrahydrofolate Selenohomocysteine MTHFR GART "5,10-Methylenetetrahydrofolate" "5,10-Methenyltetrahydrofolate" DMGDH SHMT2 PSAT1O-Phospho-4-hydroxy-L-threonine IL4I1 MTHFS Cystathionine AMT Sarcosine SARDH SHMT1 5-Formyltetrahydrofolate O-Phospho-L-serine CBS FTCD N-Formimino-L-glutamate PSPH 2-Oxo-3-hydroxy-4-phosphobutanoate NAGS GNMT NH3 L-Serine L-Pipecolate PIPOX 5-Formiminotetrahydrofolate PPIG L-Cystathionine Thiocysteine Glycine Background and hypothesis GCAT Oxaloacetate L-AlanineL-Glutamate CTH 2-Oxobutanoate L-2-Amino-3-oxobutanoic acid L-Cysteine AGXT Hydroxypyruvate SDS ANPEP 2-Aminoacrylate ALAS2 AGXT2 S-Aminomethyldihydrolipoylprotein GLDC L-Cystine ALAS1 CP GOT1 GPT GPT2 GOT2 GSS ADC CO2 5-Aminolevulinate
  • GOT2 Antibody Cat

    GOT2 Antibody Cat

    GOT2 Antibody Cat. No.: 25-873 GOT2 Antibody Specifications HOST SPECIES: Rabbit SPECIES REACTIVITY: Human, Mouse, Rat Antibody produced in rabbits immunized with a synthetic peptide corresponding a region IMMUNOGEN: of human GOT2. TESTED APPLICATIONS: ELISA, WB GOT2 antibody can be used for detection of GOT2 by ELISA at 1:312500. GOT2 antibody APPLICATIONS: can be used for detection of GOT2 by western blot at 1 μg/mL, and HRP conjugated secondary antibody should be diluted 1:50,000 - 100,000. POSITIVE CONTROL: 1) 293T Cell Lysate PREDICTED MOLECULAR 45 kDa WEIGHT: Properties PURIFICATION: Antibody is purified by peptide affinity chromatography method. CLONALITY: Polyclonal CONJUGATE: Unconjugated PHYSICAL STATE: Liquid September 29, 2021 1 https://www.prosci-inc.com/got2-antibody-25-873.html Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% BUFFER: sucrose. CONCENTRATION: batch dependent For short periods of storage (days) store at 4˚C. For longer periods of storage, store GOT2 STORAGE CONDITIONS: antibody at -20˚C. As with any antibody avoid repeat freeze-thaw cycles. Additional Info OFFICIAL SYMBOL: GOT2 ALTERNATE NAMES: GOT2, FLJ40994, kat4, kativ, mitaat, KAT4, KATIV, mitAAT ACCESSION NO.: NP_002071 PROTEIN GI NO.: 73486658 GENE ID: 2806 USER NOTE: Optimal dilutions for each application to be determined by the researcher. Background and References Glutamic-oxaloacetic transaminase is a pyridoxal phosphate-dependent enzyme which exists in cytoplasmic and inner-membrane mitochondrial forms, GOT1 and GOT2, respectively. GOT plays a role in amino acid metabolism and the urea and tricarboxylic acid cycles. The two enzymes are homodimeric and show close homology.Glutamic- oxaloacetic transaminase is a pyridoxal phosphate-dependent enzyme which exists in BACKGROUND: cytoplasmic and inner-membrane mitochondrial forms, GOT1 and GOT2, respectively.
  • The Mt Halimun-Salak Malaise Trap Project - Releasing the Most Species Rich DNA Barcode Library for Indonesia

    The Mt Halimun-Salak Malaise Trap Project - Releasing the Most Species Rich DNA Barcode Library for Indonesia

    Biodiversity Data Journal 6: e29927 doi: 10.3897/BDJ.6.e29927 Research Article The Mt Halimun-Salak Malaise Trap project - releasing the most species rich DNA Barcode library for Indonesia Bruno Cancian de Araujo‡, Stefan Schmidt‡‡, Olga Schmidt , Thomas von Rintelen§, Rosichon Ubaidillah|, Michael Balke ‡ ‡ SNSB-Zoologische Staatssammlung München, Munich, Germany § Museum für Naturkunde, Leibniz-Institut für Evolutions- und Biodiversitätsforschung, Berlin, Germany | Museum Zoologicum Bogoriense, Research Center for Biology, Indonesian Institute of Sciences, Cibinong, Indonesia Corresponding author: Bruno Cancian de Araujo ([email protected]) Academic editor: Gergin Blagoev Received: 20 Sep 2018 | Accepted: 28 Nov 2018 | Published: 19 Dec 2018 Citation: Cancian de Araujo B, Schmidt S, Schmidt O, von Rintelen T, Ubaidillah R, Balke M (2018) The Mt Halimun-Salak Malaise Trap project - releasing the most species rich DNA Barcode library for Indonesia. Biodiversity Data Journal 6: e29927. https://doi.org/10.3897/BDJ.6.e29927 Abstract The Indonesian archipelago features an extraordinarily rich biota. However, the actual taxonomic inventory of the archipelago remains highly incomplete and there is hardly any significant taxonomic activity that utilises recent technological advances. The IndoBioSys project was established as a biodiversity information system aiming at, amongst other goals, creating inventories of the Indonesian entomofauna using DNA barcoding. Here, we release the first large scale assessment of the megadiverse insect groups that occur in the Mount Halimun-Salak National Park, one of the largest tropical rain-forest ecosystem in West Java, with a focus on Hymenoptera, Coleoptera, Diptera and Lepidoptera collected with Malaise traps. From September 2015 until April 2016, 34 Malaise traps were placed in different localities in the south-eastern part of the Halimun-Salak National Park.
  • Article Soup to Tree: the Phylogeny of Beetles Inferred by Mitochondrial

    Article Soup to Tree: the Phylogeny of Beetles Inferred by Mitochondrial

    Soup to Tree: The Phylogeny of Beetles Inferred by Mitochondrial Metagenomics of a Bornean Rainforest Sample Alex Crampton-Platt,*,1,2 Martijn J.T.N. Timmermans,y,1,3 Matthew L. Gimmel,4 Sujatha Narayanan Kutty,z,1 Timothy D. Cockerill,1,5 Chey Vun Khen,6 and Alfried P. Vogler1,3,* 1Department of Life Sciences, Natural History Museum, London, United Kingdom 2Department of Genetics, Evolution and Environment, Faculty of Life Sciences, University College London, London, United Kingdom 3Division of Biology, Imperial College London, Silwood Park Campus, Ascot, United Kingdom 4Department of Biology, Faculty of Education, Palacky University, Olomouc, Czech Republic 5Department of Zoology, University of Cambridge, Cambridge, United Kingdom 6Entomology Section, Forest Research Centre, Forestry Department, Sandakan, Sabah, Malaysia yPresent address: Department of Natural Sciences, Hendon Campus, Middlesex University, London, United Kingdom zPresent address: Department of Biological Sciences, National University of Singapore, Singapore. *Corresponding author: E-mail: [email protected]; [email protected]. Associate editor: Claudia Russo Abstract In spite of the growth of molecular ecology, systematics and next-generation sequencing, the discovery and analysis of diversity is not currently integrated with building the tree-of-life. Tropical arthropod ecologists are well placed to accelerate this process if all specimens obtained through mass-trapping, many of which will be new species, could be incorporated routinely into phylogeny reconstruction. Here we test a shotgun sequencing approach, whereby mitochon- drial genomes are assembled from complex ecological mixtures through mitochondrial metagenomics, and demonstrate how the approach overcomes many of the taxonomic impediments to the study of biodiversity.