DOCSLIB.ORG

Cytoplasmic Influence on the Expression of Nuclear Genes Affecting Life Span in Drosophila M Elanogaster

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Cytoplasmic Influence on the Expression of Nuclear Genes Affecting Life Span in Drosophila M Elanogaster Heredity 66 (1991) 259—264 Received 26 June 1990 Genetical Society of Great Britain Cytoplasmic influence on the expression of nuclear genes affecting life span in Drosophila m elanogaster ISAMUYONEMURA, TOMIO MOTOYAMA,* HAYATO HASEKURA & BARRY BOETTCHERt Departmentof Legal Medicine, Shinshu University School of Medicine, Matsumoto, 390 Japan, *Department of Physiology, College of Environmental Health, Azabu University, Sagami-hara, 229 Japan and tDepartment of Biological Sciences, The University of Newcastle, New South Wales, 2308 Australia Inearlier studies we have found that the difference between short and long life spans of two inbred strains of Drosophila melanogaster is controlled by nuclear major genes. The present study has revealed a cytoplasmic factor that influences the expression of the nuclear longevity genes. The factor shows a typical maternal inheritance and is considered to be an extranuclear gene, such as mitochondrial DNA (chondriome). This paper marks the discovery of two basic forms of inheritance, nuclear and extra-nuclear, in the genetics of life span of D. melanogaster. These findings suggest that further studies, including genetic engineering, on longevity and aging might enable direct manipulation of these characters. Keywords:Drosophilamelanogasrer, extranuclear genes, gene expression, life-span, nuclear major genes. Introduction The present work reports the discovery of a new genetic influence on life span of D.m. involving extra- Fromthe results obtained in a number of studies, it has nuclear genes as well as nuclear genes, Jm A and Jm X. been concluded that life span is under polygenic Thus, two fundamental modes of inheritance, nuclear control. Genetic contribution to variation of life span is and extranuclear, are shown to influence life span. not large; environmental conditions such as tempera- ture or food are major determinants. Heritability of life span has been calculated as 0.13 3—0.442 in Drosophila Materialsand methods (Maynard Smith, 1959; Tantawy & Rakha, 1964), 0.2—0.3 in nematodes (Johnson & Wood, 1982), and Twoshort-lived purebred strains of D. m. were used as 0.21—0.36 in mice (Storer, 1966). the parental stocks, P1 and P2. These were mated Goodrick (1975), however, obtained high herit- according to the combinations listed in Table 1 to ability (0.47—0.787) of life span in mice, and his calcu- produce reciprocal F1 hybrids, F2 and all backcross lations indicated that the inheritance of life span in generations. Flies were bred at 27°C on food contain- mice was controlled by a single gene. The present ing 8 per cent dry yeast and 5 per cent sugar. The dead authors have undertaken mating experiments with two were counted every day and were removed from the purebred strains of Drosophila melanogaster (D. m.) culture vials. The P1 strain was established from a wild and have demonstrated that major genes at two loci, type stock of D.m. through 13 or more generations of autosomal and X chromosomal, determined their life sib-mating; their genes -are presumed to be homo- spans. These longevity genes were analysed for their zygous at 95 per cent or more loci. This strain is identi- mode of inheritance and were named Jm (Yonemura cal with the P1 generation (the short-lived parent) used et a!., 1989, 1990). Luckinbill et al. (1988) also in previous work (Yonemura et al., 1989, 1990). The succeeded in selection of life span in D. m., and demon- P2 strain in the present work is a purebred line estab- strated that the difference in life span in their stocks lished by the same procedure from Oregon R. Details was mainly determined by a few genes on chromosome of culture and handling methods were the same as in an 1 (X chromosome) and chromosome 3 (autosome). earlier report (Yonemura et al., 1989). 259 260 I. YONEMURA ETAL. These results show that the inheritance of life span in Results this experiment was controlled by few major genes. Themeans and standard deviations of the parental On the other hand, the scaling test of Mather (1949) stocks, the F1 and F2 generations, and the reciprocal requires the following calculations, taking the means of backcross generations are given in Table 2. These data B1, B2 and F2 generations as ,B2and F2 and those of — — ignore the obvious differences between the sexes or P1, P2 and F1 as P1, P2 and F1: A =2B1 P1 Fl, between different crosses in the same generations. B=2B2—P2--F1, C=4F2—2F1—P1—P2. However, the procedure is necessary to enable the If A =B=C=0, genes in the model are considered following calculations. to have only additive and dominance effects and to If we take the phenotypic variance in the F2 as Vi,, show no epistatic effect; epistatic effect is an interac- the genotypic variance as VG, and the environmental tion between gene loci. Calculations based on the data variance as VE, then V =V+ VE. VE is given by in Table 2 result in A=9.4±0.814, B3.4±0.804 (V+ Vp2+2VF)/4 using variances of P1, P2 and F1. and C 12.8 2.12, which are significantly larger than From Table 2 it can be calculated that V,= 289.00 and 0 (P< 0.05). This shows interaction variance is large. VE= 153.05. Heritability in the broad sense, h2b, is This is inconsistent with the above conclusion that VG given by V0/V. Then h2b can be calculated as solely consists of VA. Consequently, the quantitative h2b=0.4704. VG is divided into additive variance genetic analysis is invalid. (VA), dominance variance (VD) and interaction variance The distribution of life spans of the different genera- (V1): VG= VA+ V0+ V1. VA is given by 2VF2— VB, tions of D.m. studied here are shown in Fig. 1 (F, F1 which is calculated from Table 2 as VA= 305.75. This and F2 generations) and Fig. 2 (backcross generations). figure is larger than VG =135.95calculated before, The progenies showing the shortest mean life spans indicating that VD and V1 take no part in VG. In such a were P2 and generations originating from P2 females: genetic model the number of loci of the contributing F1_1, F2_1, B1_1, B2_1, B2_3, B2_4. The exception to genes can be calculated as N =R2/8VA= 0.03 (where R this general observation was B1_3. It is pointed out is the difference of mean life span between P1 and P2). that, although P2 females iii the parental generation initiated the B13 generation, the females in the back- Table 1 Notation of generations in reciprocal crosses cross (P1) contained no contribution (either nuclear or cytoplasmic) from P2 females. Mating Progeny In the progenies where the females used for breed- ing did not have a contribution from F2 females, F1 -2' P1dxp2Q 1I F22,B12, B13, B14, B22, long-lived flies were P19xP2d F12 observed. Such long-lived flies were not observed F1_1dxF1_19 F2_1 among the parental strains, including P1 which, simi- F12dF129 F22 larly, did not have a contribution from P2 females. P,dxF119 B11 P1dxF1_29 B12 P19xF1_1d B13 Discussionsand conclusions P19xF1_2d B14 P2dxF119 B21 Thestriking observation in this study is the marked P2dxF129 B22 difference in the life spans of progenies from reciprocal '2Qx1'11d B23 crosses. For example, F11 and F12; F21 and F22 P29xF12d B2_4 (Fig. 1). A difference between the mean life spans of male F1 flies from reciprocal crosses might be Table 2 Sample number in each generation and the mean accounted for by differences in genes located on the X life span. Values are means s.d. (days) and Y chromosomes of the parental stocks. However, such an explanation could not account for the differ- Generation Number Life span ence in the mean life spans of female F1 flies observed here (F1 and F1 -2'Fig.1). However, an extranuclear P1(d+9) 641 36.5±5.6 influence appears to be involved. All progenies where P2(d+9) 640 28.1±3.1 the cytoplasm of the zygote originated from the P2 F1(total) 1297 48.1 16.9 F2(total) 1303 43.4± 17.0 parental strain, F2, F1 —1' F2 —1' B1 —1' B2 —1' B2 , showeda unimodal, short, life span. On the other B1(total) 2539 47.0± 15.8 B24' B2(total) 2582 39.8 16.3 hand, progenies where the cytoplasm of the zygote B(total) 5121 43.4± 16.5 originated from the P1 parental strain, but the nuclear genetic material contained a contribution from the P2 CYTOPLASMIC INFLUENCE ON LIFE SPAN IN DROSOPHILA 261 A IA IX IY parental strain, F1_2 F2_2, B1_2, B1_3, B1_4, B2_2, AIAIXIXIEl showed a wide range of life spans, including long life spans not observed in the parental stocks. Consequently, the results indicate that P2 possesses a cytoplasmic factor which has a life-shortening effect which can over-ride the nuclear longevity genes, and which follows maternal inheritance accurately. This is the reason why the variance in the F1 (VF =VF + VF2) increased, and why the above quantitative genetic analysis is invalid. A2A2X2X2E2 In generations where P2 females made no contribu- tion to the cytoplasm of the zygote, longevity graphs appeared nearly identical with those in the previous reports and their genotypic values could be likewise assigned (Yonemura etal.,1989, 1990). The nuclear genes which influenced life span to lengths far greater than those of the parental stocks, are concluded as being from the P2 parents, in which their effects were countered by the life-shortening cytoplasmic factor. In order to account for the life spans of the flies observed 20 a, A2X2YlE2 here, we have to add the cytoplasmic factor, or extra- 15 AIA2XIX2E2 nuclear gene, to the Jm longevity gene systems 0 pr323 observed and studied in our previous reports (Yone- 010 - mura et a!., 1989, 1990).
Load more

Recommended publications
  • A Chloroplast Gene Is Converted Into a Nucleargene
    Proc. Nati. Acad. Sci. USA Vol. 85, pp. 391-395, January 1988 Biochemistry Relocating a gene for herbicide tolerance: A chloroplast gene is converted into a nuclear gene (QB protein/atrazine tolerance/transit peptide) ALICE Y. CHEUNG*, LAWRENCE BOGORAD*, MARC VAN MONTAGUt, AND JEFF SCHELLt: *Department of Cellular and Developmental Biology, 16 Divinity Avenue, The Biological Laboratories, Harvard University, Cambridge, MA 02138; tLaboratorium voor Genetica, Rijksuniversiteit Ghent, B-9000 Ghent, Belgium; and TMax-Planck-Institut fur Zuchtungsforschung, D-500 Cologne 30, Federal Republic of Germany Contributed by Lawrence Bogorad, September 30, 1987 ABSTRACT The chloroplast gene psbA codes for the the gene for ribulose bisphosphate carboxylase/oxygenase photosynthetic quinone-binding membrane protein Q which can transport the protein product into chloroplasts (5). We is the target of the herbicide atrazine. This gene has been have spliced the coding region of the psbA gene isolated converted into a nuclear gene. The psbA gene from an from the chloroplast DNA of the atrazine-resistant biotype atrazine-resistant biotype of Amaranthus hybridus has been of Amaranthus to the transcriptional-control and transit- modified by fusing its coding region to transcription- peptide-encoding regions of a nuclear gene, ss3.6, for the regulation and transit-peptide-encoding sequences of a bona SSU of ribulose bisphosphate carboxylase/oxygenase of pea fide nuclear gene. The constructs were introduced into the (6). The fusion-gene constructions (designated SSU-ATR) nuclear genome of tobacco by using the Agrobacteium tumor- were introduced into tobacco plants via the Agrobacterium inducing (Ti) plasmid system, and the protein product of tumor-inducing (Ti) plasmid transformation system using the nuclear psbA has been identified in the photosynthetic mem- disarmed Ti plasmid vector pGV3850 (7).
    [Show full text]
  • The Molecular Basis of Cytoplasmic Male Sterility and Fertility Restoration Patrick S
    trends in plant science reviews 24 Grandmougin, A. et al. (1989) Cyclopropyl sterols and phospholipid 34 Gachotte, D., Meens, R. and Benveniste, P. (1995) An Arabodopsis mutant composition of membrane fractions from maize roots treated with deficient in sterol biosynthesis: heterologous complementation by ERG3 fenpropimorph, Plant Physiol. 90, 591–597 encoding a ⌬7-sterol-C5-desaturase from yeast, Plant J. 8, 407–416 25 Schuler, I. et al. (1990) Soybean phosphatidylcholine vesicles containing 35 Schaller, H. et al. (1995) Expression of the Hevea brasiliensis (H.B.K.) Müll. plant sterols: a fluorescence anisotropy study, Biochim. Biophys. Acta 1028, Arg. 3-hydroxy-3-methylglutaryl coenzyme A reductase 1 in tobacco results in 82–88 sterol overproduction, Plant Physiol. 109, 761–770 26 Schuler, I. et al. (1991) Differential effects of plant sterols on water 36 Marsan, M.P., Muller, I. and Milon, A. (1996) Ability of clionasterol and permeability and on acyl chain ordering of soybean phosphatidylcholine poriferasterol (24-epimers of sitosterol and stigmasterol) to regulate membrane bilayers, Proc. Natl. Acad. Sci. U. S. A. 88, 6926–6930 lipid dynamics, Chem. Phys. Lipids 84, 117–121 27 Krajewsky-Bertrand, M-A., Milon, A. and Hartmann, M-A. (1992) 37 Goad, L.J. (1990) Application of sterol synthesis inhibitors to investigate the Deuterium-NMR investigation of plant sterol effects on soybean sterol requirements of protozoa and plants, Biochem. Soc. Trans. 18, 63–65 phosphatidylcholine acyl chain ordering, Chem. Phys. Lipids 63, 235–241 38 Cerana, R. et al. (1984) Regulating effects of brassinosteroids and of sterols 28 Grandmougin-Ferjani, A., Schuler-Muller, I.
    [Show full text]
  • “Subfossil” Koala Lemur Megaladapis Edwardsi
    Evolutionary and phylogenetic insights from a nuclear genome sequence of the extinct, giant, “subfossil” koala lemur Megaladapis edwardsi Stephanie Marciniaka, Mehreen R. Mughalb, Laurie R. Godfreyc, Richard J. Bankoffa, Heritiana Randrianatoandroa,d, Brooke E. Crowleye,f, Christina M. Bergeya,g,h, Kathleen M. Muldooni, Jeannot Randrianasyd, Brigitte M. Raharivololonad, Stephan C. Schusterj, Ripan S. Malhik,l, Anne D. Yoderm,n, Edward E. Louis Jro,1, Logan Kistlerp,1, and George H. Perrya,b,g,q,1 aDepartment of Anthropology, Pennsylvania State University, University Park, PA 16802; bBioinformatics and Genomics Intercollege Graduate Program, Pennsylvania State University, University Park, PA 16082; cDepartment of Anthropology, University of Massachusetts, Amherst, MA 01003; dMention Anthropobiologie et Développement Durable, Faculté des Sciences, Université d’Antananarivo, Antananarivo 101, Madagascar; eDepartment of Geology, University of Cincinnati, Cincinnati, OH 45220; fDepartment of Anthropology, University of Cincinnati, Cincinnati, OH 45220; gDepartment of Biology, Pennsylvania State University, University Park, PA 16802; hDepartment of Genetics, Rutgers University, New Brunswick, NJ 08854; iDepartment of Anatomy, Midwestern University, Glendale, AZ 85308; jSingapore Centre for Environmental Life Sciences Engineering, Nanyang Technological University, Singapore 639798; kDepartment of Anthropology, University of Illinois Urbana–Champaign, Urbana, IL 61801; lDepartment of Ecology, Evolution and Behavior, Carl R. Woese Institute for
    [Show full text]
  • DNA Interstrand Cross-Links Induced by Psoralen Are Not Repaired in Mammalian Mitochondria
    [CANCER RESEARCH 58. 1400-1404, April 1. 1998) Advances in Brief DNA Interstrand Cross-Links Induced by Psoralen Are Not Repaired in Mammalian Mitochondria Carleen Cullinane and Vilhelm A. Bohr1 Department of Biochemistry, Lit Trohe University, Bundoora, Victoria, 3083, Australia 1C. C./; and Laboratory of Molecular Genetics, National Institutes on Aging. Baltimore. Maryland 21224 ¡C.C., V.A. B.] Abstract evidence suggests that mitochondria are capable of repairing some types of DNA lesions. DNA damage induced by A'-methyl purines, Although it is generally known that mitochondria are defective in DNA including streptozotocin and /V-methyl-yV-nitrosourea, which, in nu damage processing, little is known about the DNA repair pathways and clear DNA, are substrates for the base excision repair pathway, are mechanisms that exist in these vital organdíes. Certain lesions that are repaired in mitochondria (3). The efficient in vivo removal of O6- removed by base excision repair are efficiently removed in mitochondria, ethyl-guanine lesions induced by ethyl nitrosourea in rat mitochondria whereas some bulky lesions that are removed by nucleotide excision repair are not repaired in these organelles. There has been much interest in has also been clearly demonstrated (4). Oxidative DNA lesions in whether mitochondria possess activities for recombination repair, and duced by alloxan (5) and bleomycin (6) are also efficiently repaired in some previous studies have reported such activities, whereas others have mitochondria. In contrast, bulky lesions induced by UVC, nitrogen not. We have taken the approach of studying the formation and removal mustard, and cisplatin are apparently not repaired by mitochondria of ¡nterstrand cross-links (ICLs) in DNA.
    [Show full text]
  • The Miniaturized Nuclear Genome of a Eukaryotic Endosymbiont Contains
    Proc. Natl. Acad. Sci. USA Vol. 93, pp. 7737-7742, July 1996 Evolution The miniaturized nuclear genome of a eukaryotic endosymbiont contains genes that overlap, genes that are cotranscribed, and the smallest known spliceosomal introns (eukaryotic operons/S13/S4/small nuclear RNP E/clp protease) PAUL R. GILSON* AND GEOFFREY I. MCFADDEN Plant Cell Biology Research Centre, School of Botany, University of Melbourne, Parkville, 3052 Victoria, Australia Communicated by Adrienne E. Clarke, University of Melbourne, Parkville, Victoria, Austrialia, February 12, 1996 (received for review December 1, 1995) ABSTRACT Chlorarachniophyte algae contain a com- (8). The nucleomorph telomere motif (TCTAGGGn) is dif- plex, multi-membraned chloroplast derived from the endo- ferent to that of the host nucleus chromosomes (TTAGGGn) symbiosis of a eukaryotic alga. The vestigial nucleus of the (8), which is consiStent with the nucleomorph being the endosymbiont, called the nucleomorph, contains only three genome of a phylogenetically unrelated endosymbiont. small linear chromosomes with a haploid genome size of 380 Previously, chlorarachniophyte nucleomorph DNA has only kb and is the smallest known eukaryotic genome. Nucleotide been shown to encode eukaryotic rRNAs that are incorpo- sequence data from a subtelomeric fragment of chromosome rated into the ribosomes in the vestigial cytoplasm surrounding III were analyzed as a preliminary investigation of the coding the nucleomorph (2). We earlier speculated the nucleomorph's capacity of this vestigial genome. Several housekeeping genes raison d'etre is to provide proteins for the maintenance of the including U6 small nuclear RNA (snRNA), ribosomal proteins chloroplast (2, 7). To synthesize these chloroplast proteins, the S4 and S13, a core protein of the spliceosome [small nuclear nucleomorph may also have to maintain genes that encode ribonucleoprotein (snRNP) E], and a clp-like protease (clpP) expression, translation and self-replication machinery (2, 7).
    [Show full text]
  • Molecular and Genetic Characterization of Rf2, A
    Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 2001 Molecular and genetic characterization of rf2, a mitochondrial aldehyde dehydrogenase gene required for male fertility in maize (Zea mays L) Xiangqin Cui Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Genetics Commons, Molecular Biology Commons, and the Plant Sciences Commons Recommended Citation Cui, Xiangqin, "Molecular and genetic characterization of rf2, a mitochondrial aldehyde dehydrogenase gene required for male fertility in maize (Zea mays L) " (2001). Retrospective Theses and Dissertations. 1037. https://lib.dr.iastate.edu/rtd/1037 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion.
    [Show full text]
  • An Alu-Based Phylogeny of Lemurs (Infraorder: Lemuriformes)
    An Alu-Based Phylogeny of Lemurs (Infraorder: Lemuriformes) Adam T. McLain1, Thomas J. Meyer1,2, Christopher Faulk1,3, Scott W. Herke1, J. Michael Oldenburg1, Matthew G. Bourgeois1, Camille F. Abshire1,4, Christian Roos5., Mark A. Batzer1*. 1 Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana, United States of America, 2 Department of Behavioral Neuroscience, Oregon Health & Science University, Portland, Oregon, United States of America, 3 Department of Environmental Health Sciences, University of Michigan, Ann Arbor, Michigan, United States of America, 4 Department of Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, Louisiana, United States of America, 5 Gene Bank of Primates and Primate Genetics Laboratory, German Primate Center, Go¨ttingen, Germany Abstract Lemurs (infraorder: Lemuriformes) are a radiation of strepsirrhine primates endemic to the island of Madagascar. As of 2012, 101 lemur species, divided among five families, have been described. Genetic and morphological evidence indicates all species are descended from a common ancestor that arrived in Madagascar ,55–60 million years ago (mya). Phylogenetic relationships in this species-rich infraorder have been the subject of debate. Here we use Alu elements, a family of primate- specific Short INterspersed Elements (SINEs), to construct a phylogeny of infraorder Lemuriformes. Alu elements are particularly useful SINEs for the purpose of phylogeny reconstruction because they are identical by descent and confounding events between loci are easily resolved by sequencing. The genome of the grey mouse lemur (Microcebus murinus) was computationally assayed for synapomorphic Alu elements. Those that were identified as Lemuriformes-specific were analyzed against other available primate genomes for orthologous sequence in which to design primers for PCR (polymerase chain reaction) verification.
    [Show full text]
  • Discordant Mitochondrial and Nuclear Gene Phylogenies in Emydid Turtles: Implications for Speciation and Conservation
    Biological Journal of the Linnean Society, 2010, 99, 445–461. With 3 figures Discordant mitochondrial and nuclear gene phylogenies in emydid turtles: implications for speciation and conservation JOHN J. WIENS1*, CAITLIN A. KUCZYNSKI1 and PATRICK R. STEPHENS2 1Department of Ecology and Evolution, Stony Brook University, Stony Brook, NY 11794-5245, USA 2Odum School of Ecology, University of Georgia, Athens, GA 30602, USA Received 1 June 2009; accepted for publication 4 August 2009bij_1342 445..461 Do phylogenies and branch lengths based on mitochondrial DNA (mtDNA) provide a reasonable approximation to those based on multiple nuclear loci? In the present study, we show widespread discordance between phylogenies based on mtDNA (two genes) and nuclear DNA (nucDNA; six loci) in a phylogenetic analysis of the turtle family Emydidae. We also find an unusual type of discordance involving the unexpected homogeneity of mtDNA sequences across species within genera. Of the 36 clades in the combined nucDNA phylogeny, 24 are contradicted by the mtDNA phylogeny, and six are strongly contested by each data set. Two genera (Graptemys, Pseudemys) show remarkably low mtDNA divergence among species, whereas the combined nuclear data show deep divergences and (for Pseudemys) strongly supported clades. These latter results suggest that the mitochondrial data alone are highly misleading about the rate of speciation in these genera and also about the species status of endangered Graptemys and Pseudemys species. In addition, despite a strongly supported phylogeny from the combined nuclear genes, we find extensive discordance between this tree and individual nuclear gene trees. Overall, the results obtained illustrate the potential dangers of making inferences about phylogeny, speciation, divergence times, and conservation from mtDNA data alone (or even from single nuclear genes), and suggest the benefits of using large numbers of unlinked nuclear loci.
    [Show full text]
  • Nuclear Gene Transformation in a Dinoflagellate
    bioRxiv preprint doi: https://doi.org/10.1101/602821; this version posted April 9, 2019. 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-NC 4.0 International license. Nuclear gene transformation in a dinoflagellate Brittany N. Sprecher, Huan Zhang*, Senjie Lin* Department of Marine Sciences, University of Connecticut, 1080 Shennecossett Rd, Groton, CT 06340 * Address for correspondence; emails [email protected]; [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/602821; this version posted April 9, 2019. 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-NC 4.0 International license. ABSTRACT The lack of a robust gene transformation tool that allows functional testing of the vast number of nuclear genes in dinoflagellates has greatly hampered our understanding of fundamental biology in this ecologically important and evolutionarily unique lineage. Here we report the development of a dinoflagellate expression vector, an electroporation protocol, and successful expression of introduced genes in the dinoflagellate Oxyrrhis marina. This protocol, involving the use of Lonza’s Nucleofector and a codon optimized antibiotic resistance gene, has been successfully used to produce consistent results in several independent experiments. It is anticipated that this protocol will be adaptable for other dinoflagellates and will allow characterization of many novel dinoflagellate genes.
    [Show full text]
  • Nuclear–Chloroplast Signalling Aravind Somanchi* and Stephen P Mayfield†
    pb2510.qxd 10/27/1999 11:58 AM Page 404 404 Nuclear–chloroplast signalling Aravind Somanchi* and Stephen P Mayfield† Chloroplast development and function relies both on structural has added new insights to the roles played by nuclear and on regulatory factors encoded within the nucleus. Recent encoded factors in controlling chloroplast functions. This work has lead to the identification of several nuclear encoded review will focus on the processes of chloroplast develop- genes that participate in a wide array of chloroplast functions. ment and differentiation, and on plastid protein expression Characterization of these genes has increased our understanding and targeting, highlighting several regulatory aspects of the of the signalling between these two compartments. Accumulating interaction between the nucleus and chloroplast involved evidence shows that a variety of molecular mechanisms are used in these key processes. for intercompartmental communication and for regulating co- ordinated chloroplast protein expression. Plastid gene expression Expression of chloroplast proteins is primarily regulated Addresses post-transcriptionally. A number of nuclear encoded factors Department of Cell Biology, The Scripps Research Institute, have been isolated that are required for plastid gene 10550 N Torrey Pines Road, La Jolla, CA 92037, USA expression. We will discuss the identification of specific *e-mail: [email protected] genes and the roles they play in plastid gene expression. †e-mail: [email protected] Current Opinion in Plant Biology 1999, 2:404–409 Transcriptional activation of plastid genes 1369-5266/99/$ — see front matter © 1999 Elsevier Science Ltd. Transcription in the chloroplast resembles that of prokary- All rights reserved. otes, particularly in the use of consensus promoter elements.
    [Show full text]
  • Nuclear Gene Dosage Effects Upon the Expression of Maize Mitochondrial Genes
    Copyright 2001 by the Genetics Society of America Nuclear Gene Dosage Effects Upon the Expression of Maize Mitochondrial Genes Donald L. Auger, Kathleen J. Newton and James A. Birchler Division of Biological Sciences, University of Missouri, Columbia, Missouri 65211 Manuscript received October 6, 2000 Accepted for publication December 21, 2000 ABSTRACT Each mitochondrion possesses a genome that encodes some of its own components. The nucleus encodes most of the mitochondrial proteins, including the polymerases and factors that regulate the expression of mitochondrial genes. Little is known about the number or location of these nuclear factors. B-A translocations were used to create dosage series for 14 different chromosome arms in maize plants with normal cytoplasm. The presence of one or more regulatory factors on a chromosome arm was indicated when variation of its dosage resulted in the alteration in the amount of a mitochondrial transcript. We used quantitative Northern analysis to assay the transcript levels of three mitochondrially encoded components of the cytochrome c oxidase complex (cox1, cox2, and cox3). Data for a nuclearly encoded component (cox5b) and for two mitochondrial genes that are unrelated to cytochrome c oxidase, ATP synthase ␣-subunit and 18S rRNA, were also determined. Two tissues, embryo and endosperm, were compared and most effects were found to be tissue speci®c. Signi®cantly, the array of dosage effects upon mitochondrial genes was similar to what had been previously found for nuclear genes. These results support the concept that although mitochondrial genes are prokaryotic in origin, their regulation has been extensively integrated into the eukaryotic cell. ITOCHONDRIA are the cellular sites for many dance of CMS transcripts (reviewed by Schnable and M energy conversion processes.
    [Show full text]
  • Mitochondrial Genetics Regulate Nuclear Gene Expression Through Metabolites COMMENTARY Jessica L
    COMMENTARY Mitochondrial genetics regulate nuclear gene expression through metabolites COMMENTARY Jessica L. Fettermana,b and Scott W. Ballingerc,1 Mitochondria contain multiple copies of mitochondrial (αKG) levels. For example, Kopinski et al. find that, under DNA (mtDNA), which encode genes essential for conditions of high heteroplasmy (A3243G), acetyl-CoA cellular bioenergetics. When more than one type of levels decrease, which associates with decreased his- mtDNA genome exists within the mitochondrion, or be- tone H4 acetylation (Fig. 1). Overall, they find that tween mitochondria, a condition termed heteroplasmy mitochondrial-derived metabolites correlate with occurs. In this respect, it has been long observed that histone posttranslational modifications, which differ differences in mtDNA heteroplasmy involving path- across the different levels of heteroplasmy. Cybrids ogenic mtDNA mutations generate a broad range with 70 to 100% A3243G heteroplasmy have lower of clinical phenotypes. For instance, it is known levels of acetyl-CoA that is associated with lower his- that increasing levels of the transfer RNA leucine tone H4 acetylation, and additionally generate less [tRNALeu(UUR)] 3243A > G mutant “result successively acetyl-CoA from glucose, instead producing higher in diabetes, neuromuscular degenerative disease, amounts of lactate, a phenotype recapitulated by and perinatal lethality” (1); however, the specific mo- inhibiting mitochondrial protein synthesis with chlor- lecular mechanisms driving these diverse clinical phe- amphenicol or complex I inhibition with rotenone. notypes have been not clearly understood. In PNAS, Additionally, cybrids with 30 to 70% A3243G have Kopinski et al. (1) advance our understanding of higher levels of αKG/succinate, which is associated this mystery by manipulating the levels of mtDNA with lower levels of histone 3 methylation (Fig.
    [Show full text]