Analysis of the Coding Sequences of Clownfish Reveals Molecular

Total Page:16

File Type:pdf, Size:1020Kb

Analysis of the Coding Sequences of Clownfish Reveals Molecular Sahm et al. BMC Evolutionary Biology (2019) 19:89 https://doi.org/10.1186/s12862-019-1409-0 RESEARCH ARTICLE Open Access Analysis of the coding sequences of clownfish reveals molecular convergence in the evolution of lifespan Arne Sahm1, Pedro Almaida-Pagán2, Martin Bens1, Mirko Mutalipassi3, Alejandro Lucas-Sánchez2, Jorge de Costa Ruiz2, Matthias Görlach1 and Alessandro Cellerino1,4* Abstract Background: Standard evolutionary theories of aging postulate that reduced extrinsic mortality leads to evolution of longevity. Clownfishes of the genus Amphiprion live in a symbiotic relationship with sea anemones that provide protection from predators. We performed a survey and identified at least two species with a lifespan of over 20 years. Given their small size and ease of captive reproduction, clownfish lend themselves as experimental models of exceptional longevity. To identify genetic correlates of exceptional longevity, we sequenced the transcriptomes of Amphiprion percula and A. clarkii and performed a scan for positively-selected genes (PSGs). Results: The PSGs that we identified in the last common clownfish ancestor were compared with PSGs detected in long-lived mole rats and short-lived killifishes revealing convergent evolution in processes such as mitochondrial biogenesis. Among individual genes, the Mitochondrial Transcription Termination Factor 1 (MTERF1), was positively- selected in all three clades, whereas the Glutathione S-Transferase Kappa 1 (GSTK1) was under positive selection in two independent clades. For the latter, homology modelling strongly suggested that positive selection targeted enzymatically important residues. Conclusions: These results indicate that specific pathways were recruited in independent lineages evolving an exceptionally extended or shortened lifespan and point to mito-nuclear balance as a key factor. Keywords: Amphiprion, Positive selection, Evolution of lifespan, Life-history trait, Mito-nuclear balance Background positive selection. A frequently used method for detecting The lifespan of vertebrate species spans more than two positive selection relies on the comparison of the orders of magnitude, from a few months for annual killi- sequence of protein-coding genes in related clades where fishes [1] to several centuries for the Greenland shark one of the clades evolved the trait of interest, in this [2]. Understanding the genetic architecture underlying specific case exceptional lifespan. To date, several different these differences is a major challenge that may deliver mammalian taxa/clades were analysed with this approach new insights into the mechanisms controlling evolution with the purpose of identifying sequence changes associ- of lifespan and human longevity. ated to evolution of longevity: the elephant, the bowhead Next-generation sequencing technology can provide whale, bats and mole-rats [3–8]. In addition to the analysis genome-scale sequence information for a large number of positive selection, other phylogeny-based methods have of species and has revolutionized evolutionary genomics. been used to investigate genetic changes related to the A particularly useful approach to identify the genetic evolution of longevity, e.g. [9–12]. The different analyses architecture of evolutionary novelties is the analysis of delivered interesting candidate genes and pathways that underwent accelerated molecular evolution in coincidence with evolution of exceptional lifespan. A major drawback * Correspondence: [email protected] of all these approaches – including the analysis of positive 1Leibniz Institute on Aging, Fritz Lipmann Institute, Jena, Germany 4Bio@SNS, Scuola Normale Superiore, Pisa, Italy selection – is that long-lived mammals are difficult or Full list of author information is available at the end of the article © The Author(s). 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Sahm et al. BMC Evolutionary Biology (2019) 19:89 Page 2 of 12 impossible to be kept in captivity and manipulated experi- are subject to selective breeding to fix specific pigmenta- mentally. This creates the need for a long-lived vertebrate tion patterns so that a number of different captive strains that is small in size, easily adaptable to captive life, can be are available. For these reasons, the clownfish could be- bred in large numbers and therefore could become a con- come the first experimental model for long-lived venient experimental model organism. vertebrates. The standard evolutionary theories of aging – namely In order to identify the genetic basis of adaptations the Mutation Accumulation [13], Antagonistic Pleiotropy linked to the clownfishes’ exceptional lifespan, we per- [14] and Disposable Soma theory [15] – predict that low formed a positive selection analysis. This analysis extrinsic mortality conditions lead to the evolution of slow requires the identification of the closest related taxon senescence and increased lifespan. Some examples that that does not possess the trait of interest in order to confirm these theories are the exceptional longevity of exclude events of positive selection that predate the evo- vertebrate species under low predation risk since they are lution of this trait [38]. chemically protected [16, 17], adapted to an arboreal life Other species of damselfish evolved an inter-specific mu- [18] or found in protected environments such as caves tualistic relationship with branching corals [39, 40]. In this [19], respectively. On the other hand, annual fishes of the case, corals are used by fishes as shelter that can provide genus Nothobranchius provide an example of how protection from predators and a safe area for egg laying increased extrinsic mortality conditions lead to the [41, 42]. Among the family Pomacentridae, Chromis viridis evolution of accelerated senescence and a short lifespan shows an interesting relationship with a wide range of scler- [20–22]. Analysis of positive selection in annual killifishes actinians [43, 44]. Despite the presence of a favourable revealed a potential link between the evolution of genes microhabitat, C. viridis arepredatedbyawiderangeofgen- governing mitochondrial biogenesis and the evolution of eralist predator species. Hixon and Carr [45]suggested lifespan [23]. there is a clear relationship among transient and benthic All clownfish species (genus Amphiprion) evolved a predators and damselfish mortality: damselfishes that specific adaptation that allows them to live in symbiosis search for protection in the shelter from transient predators with sea anemones. Symbiosis evolved in the last com- are susceptible to attack by resident benthic predators and mon ancestor of the clownfish and clownfish represent a vice versa. In the presence of both groups of predators, monophyletic group in the Pomacentridae family (dam- mortality increases dramatically due to the lack of available selfishes) [24]. In the Indo-Pacific Ocean, clownfishes refuge that expose Chromis to intense predation [45]. are found in association with one or more sea anemone Therefore, Chromis viridis represents a well-suited out- species and a large variation in host usage exists [25–27]. group for our analysis because it shares with clownfish sev- Fish that feel threatened by predators immediately seek eral general traits linked to benthic life and symbiosis with protection by the anemone’s tentacles; without that symbi- corals but it is subject to much higher predation rates osis, fish are readily attacked by predators [28–30]. There- (Fig. 1). fore, clownfishes are protected from predators through There are no reliable estimates of C. viridis lifespan in the presence of anemones [31]. Hence, the overall mortal- captivity. This is mainly due to the fact that these ity rate of the clownfish is low as compared to other coral animals build large schools and it is not possible to iden- reef fishes or other tropical species of Pomacentridae of tify them individually, as opposed to clownfishes that are thesamesize[28, 31–34]. normally kept as pairs and, if multiple pairs are main- All clownfishes are born as males and develop, tained in the aquarium, each pair associates to a differ- through protandrous hermaphroditism, into females: in ent symbiont making their distinction easy. For the same a colony, only the dominant pair contributes to the reason, the presence of a specific clownfish on a specific reproduction of the colony [35]. Other individuals of the anemone can be assessed in multiple years to obtain the colony are non-breeding males. Studies in the wild have rates of per annum mortality [36], which is not possible shown that natural mortality of adult clownfishes can be for C. viridis. However, it is certain that tropical very low: during the period 2011–2013, the average bi- Chromis fishes undergo severe predation in the annual mortality rate per capita varied, depending on post-settlement phase [45]. In one experiment where the the study site, between 0.18 and 0.49 for juveniles, 0.09 total number of the population
Recommended publications
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Barbatula Leoparda (Actinopterygii, Nemacheilidae), a New Endemic Species of Stone Loach of French Catalonia
    Scientific paper Barbatula leoparda (Actinopterygii, Nemacheilidae), a new endemic species of stone loach of French Catalonia by Camille GAULIARD (1), Agnès DETTAI (2), Henri PERSAT (1, 3), Philippe KEITH (1) & Gaël P.J. DENYS* (1, 4) Abstract. – This study described a new stone loach species in France, Barbatula leoparda, which is endemic to French Catalonia (Têt and Tech river drainages). Seven specimens were compared to 49 specimens of B. bar- batula (Linnaeus, 1758) and 71 specimens of B. quignardi (Băcescu-Meşter, 1967). This new species is char- acterized by the presence of blotches on the belly and the jugular area in individuals longer than 47 mm SL and by a greater interorbital distance (35.5 to 41.8% of the head length). We brought moreover the sequence of two mitochondrial markers (COI and 12S, respectively 652 and 950 bp) of the holotype, which are well distinct from all other species, for molecular identifications. This discovery is important for conservation. Résumé. – Barbatula leoparda (Actinopterigii, Nemacheilidae), une nouvelle espèce endémique de loche fran- che en Catalogne française. © SFI Submitted: 4 Jun. 2018 Cette étude décrit une nouvelle espèce de loche franche en France, Barbatula leoparda, qui est endémique Accepted: 23 Jan. 2019 Editor: G. Duhamel à la Catalogne française (bassins de la Têt et du Tech). Sept spécimens ont été comparés à 49 spécimens de B. barbatula (Linnaeus, 1758) et 71 spécimens de B. quignardi (Băcescu-Meşter, 1967). Cette nouvelle espèce est caractérisée par la présence de taches sur le ventre et dans la partie jugulaire pour les individus d’une taille supérieure à 47 mm LS et par une plus grande distance inter-orbitaire (35,5 to 41,8% de la longueur de la tête).
    [Show full text]
  • Supplementary Table S4. FGA Co-Expressed Gene List in LUAD
    Supplementary Table S4. FGA co-expressed gene list in LUAD tumors Symbol R Locus Description FGG 0.919 4q28 fibrinogen gamma chain FGL1 0.635 8p22 fibrinogen-like 1 SLC7A2 0.536 8p22 solute carrier family 7 (cationic amino acid transporter, y+ system), member 2 DUSP4 0.521 8p12-p11 dual specificity phosphatase 4 HAL 0.51 12q22-q24.1histidine ammonia-lyase PDE4D 0.499 5q12 phosphodiesterase 4D, cAMP-specific FURIN 0.497 15q26.1 furin (paired basic amino acid cleaving enzyme) CPS1 0.49 2q35 carbamoyl-phosphate synthase 1, mitochondrial TESC 0.478 12q24.22 tescalcin INHA 0.465 2q35 inhibin, alpha S100P 0.461 4p16 S100 calcium binding protein P VPS37A 0.447 8p22 vacuolar protein sorting 37 homolog A (S. cerevisiae) SLC16A14 0.447 2q36.3 solute carrier family 16, member 14 PPARGC1A 0.443 4p15.1 peroxisome proliferator-activated receptor gamma, coactivator 1 alpha SIK1 0.435 21q22.3 salt-inducible kinase 1 IRS2 0.434 13q34 insulin receptor substrate 2 RND1 0.433 12q12 Rho family GTPase 1 HGD 0.433 3q13.33 homogentisate 1,2-dioxygenase PTP4A1 0.432 6q12 protein tyrosine phosphatase type IVA, member 1 C8orf4 0.428 8p11.2 chromosome 8 open reading frame 4 DDC 0.427 7p12.2 dopa decarboxylase (aromatic L-amino acid decarboxylase) TACC2 0.427 10q26 transforming, acidic coiled-coil containing protein 2 MUC13 0.422 3q21.2 mucin 13, cell surface associated C5 0.412 9q33-q34 complement component 5 NR4A2 0.412 2q22-q23 nuclear receptor subfamily 4, group A, member 2 EYS 0.411 6q12 eyes shut homolog (Drosophila) GPX2 0.406 14q24.1 glutathione peroxidase
    [Show full text]
  • Characterization and Proteomic-Transcriptomic Investigation of Monocarboxylate Transporter 6 Knockout Mice
    Supplemental material to this article can be found at: http://molpharm.aspetjournals.org/content/suppl/2019/07/10/mol.119.116731.DC1 1521-0111/96/3/364–376$35.00 https://doi.org/10.1124/mol.119.116731 MOLECULAR PHARMACOLOGY Mol Pharmacol 96:364–376, September 2019 Copyright ª 2019 by The American Society for Pharmacology and Experimental Therapeutics Characterization and Proteomic-Transcriptomic Investigation of Monocarboxylate Transporter 6 Knockout Mice: Evidence of a Potential Role in Glucose and Lipid Metabolism s Robert S. Jones, Chengjian Tu, Ming Zhang, Jun Qu, and Marilyn E. Morris Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, New York (R.S.J., C.T., J.Q., M.E.M.); and New York State Center of Excellence in Bioinformatics and Life Sciences, Buffalo, New York (C.T., M.Z., J.Q.) Received March 21, 2019; accepted June 27, 2019 Downloaded from ABSTRACT Monocarboxylate transporter 6 [(MCT6), SLC16A5] is an or- Mct62/2 mice demonstrated significant changes in 199 genes phan transporter with no known endogenous substrates or in the liver compared with Mct61/1 mice. In silico biological physiological role. Previous in vitro and in vivo experiments pathway analyses revealed significant changes in proteins and molpharm.aspetjournals.org investigated MCT6 substrate/inhibitor specificity in Xenopus genes involved in glucose and lipid metabolism–associated laevis oocytes; however, these data remain limited. Transcrip- pathways. This study is the first to provide evidence for an tomic changes in the livers of mice undergoing different dieting association of Mct6 in the regulation of glucose and lipid schemes have suggested that Mct6 plays a role in glucose metabolism.
    [Show full text]
  • Multi-Locus Fossil-Calibrated Phylogeny of Atheriniformes (Teleostei, Ovalentaria)
    Molecular Phylogenetics and Evolution 86 (2015) 8–23 Contents lists available at ScienceDirect Molecular Phylogenetics and Evolution journal homepage: www.elsevier.com/locate/ympev Multi-locus fossil-calibrated phylogeny of Atheriniformes (Teleostei, Ovalentaria) Daniela Campanella a, Lily C. Hughes a, Peter J. Unmack b, Devin D. Bloom c, Kyle R. Piller d, ⇑ Guillermo Ortí a, a Department of Biological Sciences, The George Washington University, Washington, DC, USA b Institute for Applied Ecology, University of Canberra, Australia c Department of Biology, Willamette University, Salem, OR, USA d Department of Biological Sciences, Southeastern Louisiana University, Hammond, LA, USA article info abstract Article history: Phylogenetic relationships among families within the order Atheriniformes have been difficult to resolve Received 29 December 2014 on the basis of morphological evidence. Molecular studies so far have been fragmentary and based on a Revised 21 February 2015 small number taxa and loci. In this study, we provide a new phylogenetic hypothesis based on sequence Accepted 2 March 2015 data collected for eight molecular markers for a representative sample of 103 atheriniform species, cover- Available online 10 March 2015 ing 2/3 of the genera in this order. The phylogeny is calibrated with six carefully chosen fossil taxa to pro- vide an explicit timeframe for the diversification of this group. Our results support the subdivision of Keywords: Atheriniformes into two suborders (Atherinopsoidei and Atherinoidei), the nesting of Notocheirinae Silverside fishes within Atherinopsidae, and the monophyly of tribe Menidiini, among others. We propose taxonomic Marine to freshwater transitions Marine dispersal changes for Atherinopsoidei, but a few weakly supported nodes in our phylogeny suggests that further Molecular markers study is necessary to support a revised taxonomy of Atherinoidei.
    [Show full text]
  • Gene Ontology Functional Annotations and Pleiotropy
    Network based analysis of genetic disease associations Sarah Gilman Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy under the Executive Committee of the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2014 © 2013 Sarah Gilman All Rights Reserved ABSTRACT Network based analysis of genetic disease associations Sarah Gilman Despite extensive efforts and many promising early findings, genome-wide association studies have explained only a small fraction of the genetic factors contributing to common human diseases. There are many theories about where this “missing heritability” might lie, but increasingly the prevailing view is that common variants, the target of GWAS, are not solely responsible for susceptibility to common diseases and a substantial portion of human disease risk will be found among rare variants. Relatively new, such variants have not been subject to purifying selection, and therefore may be particularly pertinent for neuropsychiatric disorders and other diseases with greatly reduced fecundity. Recently, several researchers have made great progress towards uncovering the genetics behind autism and schizophrenia. By sequencing families, they have found hundreds of de novo variants occurring only in affected individuals, both large structural copy number variants and single nucleotide variants. Despite studying large cohorts there has been little recurrence among the genes implicated suggesting that many hundreds of genes may underlie these complex phenotypes. The question
    [Show full text]
  • Training Manual Series No.15/2018
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by CMFRI Digital Repository DBTR-H D Indian Council of Agricultural Research Ministry of Science and Technology Central Marine Fisheries Research Institute Department of Biotechnology CMFRI Training Manual Series No.15/2018 Training Manual In the frame work of the project: DBT sponsored Three Months National Training in Molecular Biology and Biotechnology for Fisheries Professionals 2015-18 Training Manual In the frame work of the project: DBT sponsored Three Months National Training in Molecular Biology and Biotechnology for Fisheries Professionals 2015-18 Training Manual This is a limited edition of the CMFRI Training Manual provided to participants of the “DBT sponsored Three Months National Training in Molecular Biology and Biotechnology for Fisheries Professionals” organized by the Marine Biotechnology Division of Central Marine Fisheries Research Institute (CMFRI), from 2nd February 2015 - 31st March 2018. Principal Investigator Dr. P. Vijayagopal Compiled & Edited by Dr. P. Vijayagopal Dr. Reynold Peter Assisted by Aditya Prabhakar Swetha Dhamodharan P V ISBN 978-93-82263-24-1 CMFRI Training Manual Series No.15/2018 Published by Dr A Gopalakrishnan Director, Central Marine Fisheries Research Institute (ICAR-CMFRI) Central Marine Fisheries Research Institute PB.No:1603, Ernakulam North P.O, Kochi-682018, India. 2 Foreword Central Marine Fisheries Research Institute (CMFRI), Kochi along with CIFE, Mumbai and CIFA, Bhubaneswar within the Indian Council of Agricultural Research (ICAR) and Department of Biotechnology of Government of India organized a series of training programs entitled “DBT sponsored Three Months National Training in Molecular Biology and Biotechnology for Fisheries Professionals”.
    [Show full text]
  • Evolution and Ecology in Widespread Acoustic Signaling Behavior Across Fishes
    bioRxiv preprint doi: https://doi.org/10.1101/2020.09.14.296335; this version posted September 14, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Evolution and Ecology in Widespread Acoustic Signaling Behavior Across Fishes 2 Aaron N. Rice1*, Stacy C. Farina2, Andrea J. Makowski3, Ingrid M. Kaatz4, Philip S. Lobel5, 3 William E. Bemis6, Andrew H. Bass3* 4 5 1. Center for Conservation Bioacoustics, Cornell Lab of Ornithology, Cornell University, 159 6 Sapsucker Woods Road, Ithaca, NY, USA 7 2. Department of Biology, Howard University, 415 College St NW, Washington, DC, USA 8 3. Department of Neurobiology and Behavior, Cornell University, 215 Tower Road, Ithaca, NY 9 USA 10 4. Stamford, CT, USA 11 5. Department of Biology, Boston University, 5 Cummington Street, Boston, MA, USA 12 6. Department of Ecology and Evolutionary Biology and Cornell University Museum of 13 Vertebrates, Cornell University, 215 Tower Road, Ithaca, NY, USA 14 15 ORCID Numbers: 16 ANR: 0000-0002-8598-9705 17 SCF: 0000-0003-2479-1268 18 WEB: 0000-0002-5669-2793 19 AHB: 0000-0002-0182-6715 20 21 *Authors for Correspondence 22 ANR: [email protected]; AHB: [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.09.14.296335; this version posted September 14, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.
    [Show full text]
  • Evolutionary History and Whole Genome Sequence of Pejerrey (Odontesthes Bonariensis): New Insights Into Sex Determination in Fishes
    Evolutionary History and Whole Genome Sequence of Pejerrey (Odontesthes bonariensis): New Insights into Sex Determination in Fishes by Daniela Campanella B.Sc. in Biology, July 2009, Universidad Nacional de La Plata, Argentina A Dissertation submitted to The Faculty of The Columbian College of Arts and Sciences of The George Washington University in partial fulfillment of the requirements for the degree of Doctor of Philosophy January 31, 2015 Dissertation co-directed by Guillermo Ortí Louis Weintraub Professor of Biology Elisabet Caler Program Director at National Heart, Lung and Blood Institute, NIH The Columbian College of Arts and Sciences of The George Washington University certifies that Daniela Campanella has passed the Final Examination for the degree of Doctor of Philosophy as of December 12th, 2014. This is the final and approved form of the dissertation. Evolutionary History and Whole Genome Sequence of Pejerrey (Odontesthes bonariensis): New Insights into Sex Determination in Fishes Daniela Campanella Dissertation Research Committee: Guillermo Ortí, Louis Weintraub Professor of Biology, Dissertation Co-Director Elisabet Caler, Program Director at National Heart, Lung and Blood Institute, NIH, Dissertation Co-Director Hernán Lorenzi, Assistant Professor in Bioinformatics Department, J. Craig Venter Institute Rockville Maryland, Committee Member Jeremy Goecks, Assistant Professor of Computational Biology, Committee Member ! ""! ! Copyright 2015 by Daniela Campanella All rights reserved ! """! Dedication The author wishes to dedicate this dissertation to: My love, Ford, for his unconditional support and inspiration. For teaching me that admiration towards each other’s work is the fundamental fuel to go anywhere. My family and friends, for being there, meaning “there” everywhere and whenever. My grandpa Hugo, a pejerrey lover who knew how to fish, cook and enjoy the “silver arrows”.
    [Show full text]
  • PRIMER NOTES Microsatellite Loci Isolated from Odontesthes
    MEC882.fm Page 629 Friday, March 31, 2000 8:44 AM Molecular Ecology (2000) 9, 629–644 PRIMER Blackwell Science, Ltd NOTES Microsatellite00Graphicraft Limited, Hong Kong loci isolated from for sequencing. This strategy increased our chances of choosing both long and short repeats as they are expected to inform at Odontesthes argentinensis and the different levels of divergence (see below). Thirteen of the clones O. perugiae species group and their use sequenced contained microsatellites with appropriate flank- in other South American silverside fish ing regions for primer design. Variation at each of the 13 loci was assessed by the poly- L. B. BEHEREGARAY and P. SUNNUCKS merase chain reaction (PCR) using a 10 µL radiolabelled reaction containing 50 –100 ng of template DNA, 12 pmol of each Department of Biological Sciences, Macquarie University, NSW 2109, primer, 0.5 units of Taq DNA polymerase (Promega), 200 µm Australia µ of dCTP, dGTP, and dTTP, 20 m of dATP, 2 mm MgCl2, 10 mm Keywords: microsatellites, Odontesthes, population genetics, radiation, Tris-HCl (pH 8.3), 50 mm KCl, 0.1% Triton X-100 and 0.05 µL silverside fish [α-33P] dATP at 1000 Ci/mmol overlaid with mineral oil. PCR Received 4 September 1999; revision received 18 October 1999; accepted amplifications were performed in an MJ Research PTC100 23 October 1999 thermocycler starting with 94 °C for 3 min, followed by a ‘touchdown’ (32 cycles at 94 °C/20 s, annealing/45 s and Correspondence: L. B. Beheregaray. Fax: (612) 9850 8245. E-mail: 72 °C/60 s), and a final step of 72 °C for 4 min.
    [Show full text]
  • (Cos) Rnas and of a Conserved Family of Organellar RNA-Binding Proteins, the Heptatricopeptide Repeat Proteins, in the Malaria Parasite Arne Hillebrand1, Joachim M
    Published online 8 August 2018 Nucleic Acids Research, 2018, Vol. 46, No. 19 10417–10431 doi: 10.1093/nar/gky710 Identification of clustered organellar short (cos) RNAs and of a conserved family of organellar RNA-binding proteins, the heptatricopeptide repeat proteins, in the malaria parasite Arne Hillebrand1, Joachim M. Matz2, Martin Almendinger1,KatjaMuller¨ 2, Kai Matuschewski2 and Christian Schmitz-Linneweber1,* 1Humboldt University Berlin, Molecular Genetics, Berlin, Germany and 2Humboldt University, Department of Molecular Parasitology, Berlin, Germany Received December 12, 2017; Revised July 20, 2018; Editorial Decision July 23, 2018; Accepted July 24, 2018 ABSTRACT malaria globally, resulting in almost half a million deaths (1). While some antimalarial treatments are available, para- Gene expression in mitochondria of Plasmodium site resistance is a continuing challenge. Plasmodium spp. falciparum is essential for parasite survival. The belong to the family of apicomplexan parasites, most of molecular mechanisms of Plasmodium organellar which contain a non-photosynthetic plastid called the api- gene expression remain poorly understood. This coplast (2). As a remnant of a red algae chloroplast, the api- includes the enigmatic assembly of the mitochon- coplast contains its own DNA, as does the Plasmodium mi- drial ribosome from highly fragmented rRNAs. Here, tochondrion. The Plasmodium nuclear genome has a size we present the identification of clustered organellar of 24 MB and contains >5000 genes; by contrast, the api- short RNA fragments (cosRNAs) that are possible coplast and mitochondrial genomes are small––35 and 6 kb, footprints of RNA-binding proteins (RBPs) in Plas- respectively. modium organelles. In plants, RBPs of the pentatri- In keeping with the descendance of the apicoplast from cyanobacteria, the apicoplast genome is organized similarly copeptide repeat (PPR) class produce footprints as a to bacterial chromosomes.
    [Show full text]
  • A SARS-Cov-2 Protein Interaction Map Reveals Targets for Drug Repurposing
    Article A SARS-CoV-2 protein interaction map reveals targets for drug repurposing https://doi.org/10.1038/s41586-020-2286-9 A list of authors and affiliations appears at the end of the paper Received: 23 March 2020 Accepted: 22 April 2020 A newly described coronavirus named severe acute respiratory syndrome Published online: 30 April 2020 coronavirus 2 (SARS-CoV-2), which is the causative agent of coronavirus disease 2019 (COVID-19), has infected over 2.3 million people, led to the death of more than Check for updates 160,000 individuals and caused worldwide social and economic disruption1,2. There are no antiviral drugs with proven clinical efcacy for the treatment of COVID-19, nor are there any vaccines that prevent infection with SARS-CoV-2, and eforts to develop drugs and vaccines are hampered by the limited knowledge of the molecular details of how SARS-CoV-2 infects cells. Here we cloned, tagged and expressed 26 of the 29 SARS-CoV-2 proteins in human cells and identifed the human proteins that physically associated with each of the SARS-CoV-2 proteins using afnity-purifcation mass spectrometry, identifying 332 high-confdence protein–protein interactions between SARS-CoV-2 and human proteins. Among these, we identify 66 druggable human proteins or host factors targeted by 69 compounds (of which, 29 drugs are approved by the US Food and Drug Administration, 12 are in clinical trials and 28 are preclinical compounds). We screened a subset of these in multiple viral assays and found two sets of pharmacological agents that displayed antiviral activity: inhibitors of mRNA translation and predicted regulators of the sigma-1 and sigma-2 receptors.
    [Show full text]