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1. Small RNA fragments derived from multiple RNA classes – the missing element of multi-omics characteristics of the hepatitis C virus cell culture model

Author

Agnieszka Zmienko, Anna Hojka-Osinska, Agata Budkowska, Marek Figlerowicz, Anna Philips, Paulina Jackowiak, Lucyna Budzko, Patrick Maillard, Institute of Bioorganic Chemistry [Poznań], Polska Akademia Nauk = Polish Academy of Sciences (PAN), Institute of Computing Science [Poznan], Poznan University of Technology (PUT), Hépacivirus et Immunité innée, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), This study was financed by the National Science Centre grant awarded on the basis of a decision number DEC-2012/05/D/NZ2/02238, to PJ, and by the Polish Ministry of Science and Higher Education grant IP2012 014972, to PJ. Partial financial support was also provided by the European Union within the European Regional Development Fund through the MPD program and by the Polish Ministry of Science and Higher Education, under the KNOW program., We thank T. Wakita for providing us (Institut Pasteur, Paris, France) with JFH-1 infectious clone and C. Rice for Huh-7.5 cells. We are indebted to the Plateforme D’Imagerie Dynamique (PFID) of the Institut Pasteur 'Imagopole' for microscopy assistance. The RNA-Seq analysis was performed using the facilities of European Centre of Bioinformatics and Genomics (Poznan, Poland)., and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Small RNA, RNA, Untranslated, genetic structures, lcsh:QH426-470, Hepatitis C virus, [SDV]Life Sciences [q-bio], lcsh:Biotechnology, Gene Dosage, Hepacivirus, Biology, medicine.disease_cause, Cell Line, Transcriptome, RNA fragments, 03 medical and health sciences, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], lcsh:TP248.13-248.65, microRNA, Genetics, medicine, Humans, tRF, Nucleic acid structure, skin and connective tissue diseases, Non-coding RNA, High-Throughput Nucleotide Sequencing, RNA, Genomics, 3. Good health, RNA silencing, lcsh:Genetics, 030104 developmental biology, HCV, RNA, Viral, Research Article, Biotechnology
Abstract

Background A pool of small RNA fragments (RFs) derived from diverse cellular RNAs has recently emerged as a rich source of functionally relevant molecules. Although their formation and accumulation has been connected to various stress conditions, the knowledge on RFs produced upon viral infections is very limited. Here, we applied the next generation sequencing (NGS) to characterize RFs generated in the hepatitis C virus (HCV) cell culture model (HCV-permissive Huh-7.5 cell line). Results We found that both infected and non-infected cells contained a wide spectrum of RFs derived from virtually all RNA classes. A significant fraction of identified RFs accumulated to similar levels as miRNAs. Our analysis, focused on RFs originating from constitutively expressed non-coding RNAs, revealed three major patterns of parental RNA cleavage. We found that HCV infection induced significant changes in the accumulation of low copy number RFs, while subtly altered the levels of high copy number ones. Finally, the candidate RFs potentially relevant for host-virus interactions were identified. Conclusions Our results indicate that RFs should be considered an important component of the Huh-7.5 transcriptome and suggest that the main factors influencing the RF biogenesis are the RNA structure and RNA protection by interacting proteins. The data presented here significantly complement the existing transcriptomic, miRnomic, proteomic and metabolomic characteristics of the HCV cell culture model. Electronic supplementary material The online version of this article (doi:10.1186/s12864-017-3891-3) contains supplementary material, which is available to authorized users.

Published
2017
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2. Metabolic and Organelle Morphology Defects in Mice and Human Patients Define Spinocerebellar Ataxia Type 7 as a Mitochondrial Disease

Author

Pawel M. Switonski, Ronald M. Evans, Fanny Mochel, Weiwei Fan, Christopher E. Wall, Chenchen Niu, Chizuru Kinoshita, Farid Ichou, Jacqueline M. Ward, Alysson R. Muotri, Albert R. La Spada, Colleen A. Stoyas, Isaac Adanyeguh, Alexandra Durr, Richard S. Morrison, Bryce L. Sopher, Brett Collins, University of California [San Diego] (UC San Diego), University of California, Duke University [Durham], Institute of Bioorganic Chemistry [Poznań], Polska Akademia Nauk = Polish Academy of Sciences (PAN), Polish Academy of Sciences (PAN), Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Institute of cardiometabolism and nutrition (ICAN), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), University of Washington [Seattle], Service de Génétique Cytogénétique et Embryologie [CHU Pitié-Salpêtrière], CHU Pitié-Salpêtrière [AP-HP], and Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)

Subjects
0301 basic medicine, Blood Glucose, Mitochondrial Diseases, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Purkinje cell, Medical Physiology, Nicotinamide adenine dinucleotide, Mitochondrion, Neurodegenerative, chemistry.chemical_compound, Purkinje Cells, Mice, 0302 clinical medicine, spinocerebellar ataxia, Neural Stem Cells, oxidative metabolism, 2.1 Biological and endogenous factors, Aetiology, lcsh:QH301-705.5, Kynurenine, Tryptophan, 3. Good health, Cell biology, Mitochondria, mitochondria, medicine.anatomical_structure, Phenotype, Adipose Tissue, Neurological, Spinocerebellar ataxia, Stem cell, trinucleotide repeat, Ataxin 7, induced pluripotent stem cells, Mitochondrial disease, mouse model, Biology, Article, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Rare Diseases, medicine, Genetics, Animals, Humans, Spinocerebellar Ataxias, Metabolomics, nicotinamide adenine dinucleotide, Organelles, Ataxin-7, Neurosciences, Reproducibility of Results, ataxin-7, medicine.disease, Stem Cell Research, NAD, Brain Disorders, 030104 developmental biology, lcsh:Biology (General), chemistry, biology.protein, NAD+ kinase, Biochemistry and Cell Biology, Peptides, Trinucleotide Repeat Expansion, Energy Metabolism, polyglutamine, 030217 neurology & neurosurgery
Abstract

SUMMARY Spinocerebellar ataxia type 7 (SCA7) is a retinal-cerebellar degenerative disorder caused by CAG-polyglutamine (polyQ) repeat expansions in the ataxin-7 gene. As many SCA7 clinical phenotypes occur in mitochondrial disorders, and magnetic resonance spectroscopy of patients revealed altered energy metabolism, we considered a role for mitochondrial dysfunction. Studies of SCA7 mice uncovered marked impairments in oxygen consumption and respiratory exchange. When we examined cerebellar Purkinje cells in mice, we observed mitochondrial network abnormalities, with enlarged mitochondria upon ultrastructural analysis. We developed stem cell models from patients and created stem cell knockout rescue systems, documenting mitochondrial morphology defects, impaired oxidative metabolism, and reduced expression of nicotinamide adenine dinucleotide (NAD+) production enzymes in SCA7 models. We observed NAD+ reductions in mitochondria of SCA7 patient NPCs using ratiometric fluorescent sensors and documented alterations in tryptophan-kynurenine metabolism in patients. Our results indicate that mitochondrial dysfunction, stemming from decreased NAD+, is a defining feature of SCA7., Graphical Abstract, In Brief Ward et al. document altered metabolism and mitochondrial dysfunction in SCA7 patients, mice, and human stem cell-derived neurons. They link these abnormalities to reduced nicotinamide adenine dinucleotide in specific subcellular compartments. Given the role of mitochondrial impairment in neurodegeneration, their results have therapeutic implications for SCA7 and related neurological disorders.

Published
2019
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3. The genome of the vervet ( Chlorocebus aethiops sabaeus )

Author

Nicoletta Archidiacono, Jennifer F. Hughes, George M. Weinstock, Yoon Jung, Christopher A. Schmitt, Angela Noll, Nelson B. Freimer, Matthew J. Jorgensen, Jennifer J. Tuscher, Nam Tran, Jason M. Brenchley, David H. O’Connor, Matthew W. Hahn, Eugene Redmond, Alex Nisbett, Roland Zahn, Hannes Svardal, Gary P. Schroth, Raquel García-Pérez, Magnus Nordborg, Nikoleta Juretic, Michael J. Montague, Gregg W.C. Thomas, Michel M. Dione, Julie A. Karl, Anna J. Jasinska, Wesley C. Warren, Jay R. Kaplan, Richard K. Wilson, Patrick Minx, Bronwen Aken, Mariano Rocchi, Colby Chiang, Françoise Thibaud-Nissen, Jessica Wasserscheid, Vasily Ramensky, Oi Wa Choi, Gennady Churakov, Roscoe Stanyon, Yu S. Huang, David Webb, Brian J. Raney, Jörn E. Schmitz, Milinn Kremitzki, Beatrice Jacquelin, Michaela Müller-Trutwin, Juergen Schmitz, Rishi Nag, Tomas Marques-Bonet, LaDeana W. Hillier, Martin Antonio, Tina Graves, Trudy R. Turner, Kim Kyung, Ken Dewar, Chad Tomlinson, Roger W. Wiseman, Oronzo Capozzi, The McDonnell Genome Institute (MGI), Washington University in Saint Louis (WUSTL), Semel Institute for Neuroscience and Human Behavior [Los Angeles, Ca], University of California [Los Angeles] (UCLA), University of California-University of California, Institute of Bioorganic Chemistry [Poznań], Polska Akademia Nauk = Polish Academy of Sciences (PAN), Institució Catalana de Recerca i Estudis Avançats (ICREA), Centro Nacional de Analisis Genomico [Barcelona] (CNAG), Gregor Mendel Institute of Molecular Plant Biology (GMI), Austrian Academy of Sciences (OeAW), University of Bari Aldo Moro (UNIBA), Whitehead Institute, Massachusetts Institute of Technology (MIT), McGill University = Université McGill [Montréal, Canada], University of Wisconsin - Milwaukee, University of the Free State [South Africa], University of Wisconsin-Madison, Beth Israel Deaconess Medical Center [Boston] (BIDMC), Harvard Medical School [Boston] (HMS), Crucell Holland B.V, St. Kitts Biomedical Research Foundation, Régulation des Infections Rétrovirales, Institut Pasteur [Paris], National Institute of Allergy and Infectious Diseases [Bethesda] (NIAID-NIH), National Institutes of Health [Bethesda] (NIH), Medical Research Council Unit The Gambia (MRC), Illumina Incorporated [San Diego, CA, USA], Wake Forest School of Medicine [Winston-Salem], Wake Forest Baptist Medical Center, Indiana University [Bloomington], Indiana University System, University of California [Santa Cruz] (UCSC), University of California, European Molecular Biology Laboratory [Hinxton], Institute of Experimental Pathology (ZMBE), Westfälische Wilhelms-Universität Münster (WWU), Institute for Evolution and Biodiversity (IEB), Università degli Studi di Firenze = University of Florence [Firenze] (UNIFI), National Center for Biotechnology Information (NCBI), The Jackson Laboratory [Bar Harbor] (JAX), Funding to R.K.W. was provided by NIH-NHGRI grant 5U54HG00307907. Support for the Vervet Research Colony was provided by NIH grant RR019963/OD010965 to J.R.K. Funding to N.B.F. was provided by NIH grants R01RR016300 and R01OD010980. The French National Agency for Research on AIDS and Viral Hepatitis (ANRS) provided funding to M.C.M.-T. Funding to M.R. and R.S. was provided by the Ministero della Universita’ e della Ricerca. Funding to K.D. was provided by Genome Canada and Genome Quebec. B.A. and R.N. acknowledge support from the Wellcome Trust (grant number WT095908) and the European Molecular Biology Laboratory., We thank the following organizations for providing sampling permits and permissions: Ethiopian Wildlife Conservation Authority, Zambia Wildlife Authority, Wildlife Division, Forestry Commission, Republic of Ghana, and Gambia Department of Parks and Wildlife Management. We thank Dr. James L. Blanchard and Dr. Ivona Pandrea for vervet samples used to characterize MHC. We thank Josh McMichael and Josh Peck for figure assistance. We thank Joanne Nelson and Barbara Gillam for data submission. We thank the members of the library production group led by Catrina Fronick and sequencing led by Matt Cordes. We also thank the Medical Research Council Unit (MRC), The Gambia for their assistance, as well as all the veterinarians who worked with us to safely obtain samples., National Institutes of Health (US), National Human Genome Research Institute (US), Agence Nationale de Recherches sur le SIDA et les Hépatites Virales (France), Agence Nationale de la Recherche (France), Ministero dell'Istruzione, dell'Università e della Ricerca, Genome Canada, EMBO, Wellcome Trust, University of California (UC)-University of California (UC), Università degli studi di Bari Aldo Moro = University of Bari Aldo Moro (UNIBA), University of the Free State [South Africa] (UFS), Institut Pasteur [Paris] (IP), University of California [Santa Cruz] (UC Santa Cruz), University of California (UC), Westfälische Wilhelms-Universität Münster = University of Münster (WWU), and Università degli Studi di Firenze = University of Florence (UniFI)

Subjects
animal diseases, [SDV]Life Sciences [q-bio], Chlorocebus aethiops, Cercopithecus aethiops, Medical and Health Sciences, Major Histocompatibility Complex, MESH: Animals, MESH: Genetic Variation, MESH: Phylogeny, Genetics (clinical), Phylogeny, MESH: Evolution, Molecular, Gene Rearrangement, education.field_of_study, Cercopithecini, Genome, biology, MESH: Genomics, [SDV.BID.EVO]Life Sciences [q-bio]/Biodiversity/Populations and Evolution [q-bio.PE], MESH: Karyotype, Genomics, Biological Sciences, 3. Good health, Rhesus macaque, Phylogeography, MESH: Phylogeography, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDE]Environmental Sciences, Complete Genome assembly, primates, cercopithecini, Infection, Biotechnology, MESH: Computational Biology, Resource, Evolution, Bioinformatics, MESH: Gene Rearrangement, Population, Karyotype, Simis, Old World monkey, MESH: Molecular Sequence Annotation, [SDV.BID]Life Sciences [q-bio]/Biodiversity, Chromosome Painting, Evolution, Molecular, MESH: Major Histocompatibility Complex, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], parasitic diseases, Genetics, Animals, MESH: Genome, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, education, [SDV.GEN]Life Sciences [q-bio]/Genetics, Genètica animal, MESH: Chromosome Painting, Human Genome, Molecular, Computational Biology, Genetic Variation, Molecular Sequence Annotation, Gene rearrangement, biology.organism_classification, MESH: Cercopithecus aethiops, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, Evolutionary biology, African Green Monkey
Abstract

Warren, Wesley C. et al., We describe a genome reference of the African green monkey or vervet (Chlorocebus aethiops). This member of the Old World monkey (OWM) superfamily is uniquely valuable for genetic investigations of simian immunodeficiency virus (SIV), for which it is the most abundant natural host species, and of a wide range of health-related phenotypes assessed in Caribbean vervets (C. a. sabaeus), whose numbers have expanded dramatically since Europeans introduced small numbers of their ancestors from West Africa during the colonial era. We use the reference to characterize the genomic relationship between vervets and other primates, the intra-generic phylogeny of vervet subspecies, and genome-wide structural variations of a pedigreed C. a. sabaeus population. Through comparative analyses with human and rhesus macaque, we characterize at high resolution the unique chromosomal fission events that differentiate the vervets and their close relatives from most other catarrhine primates, in whom karyotype is highly conserved. We also provide a summary of transposable elements and contrast these with the rhesus macaque and human. Analysis of sequenced genomes representing each of the main vervet subspecies supports previously hypothesized relationships between these populations, which range across most of sub-Saharan Africa, while uncovering high levels of genetic diversity within each. Sequence-based analyses of major histocompatibility complex (MHC) polymorphisms reveal extremely low diversity in Caribbean C. a. sabaeus vervets, compared to vervets from putatively ancestral West African regions. In the C. a. sabaeus research population, we discover the first structural variations that are, in some cases, predicted to have a deleterious effect; future studies will determine the phenotypic impact of these variations., Funding to R.K.W. was provided by NIH-NHGRI grant 5U54HG00307907. Support for the Vervet Research Colony was provided by NIH grant RR019963/OD010965 to J.R.K. Funding to N.B.F. was provided by NIH grants R01RR016300 and R01OD010980. The French National Agency for Research on AIDS and Viral Hepatitis (ANRS) provided funding to M.C.M.-T. Funding to M.R. and R.S. was provided by the Ministero della Universita’ e della Ricerca. Funding to K.D. was provided by Genome Canada and Genome Quebec. B.A. and R.N. acknowledge support from the Wellcome Trust (grant number WT095908) and the European Molecular Biology Laboratory.

Published
2015
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4. Distinctive structural motifs of RNA G-quadruplexes composed of AGG, CGG and UGG trinucleotide repeats

Author

Zofia Gdaniec, Ryszard Kierzek, Magdalena Malgowska, Dorota Gudanis, Valérie Gabelica, Eliza Wyszko, Gabelica, Valérie, Institute of Bioorganic Chemistry [Poznań], Polska Akademia Nauk = Polish Academy of Sciences (PAN), Laboratoire de spectrométrie de masse, Université de Liège-Groupe Interdisciplinaire de Génoprotéomique Appliquée-Département de Chimie, ARN : régulations naturelle et artificielle, Université Bordeaux Segalen - Bordeaux 2-Institut Européen de Chimie et de Biologie-Institut National de la Santé et de la Recherche Médicale (INSERM), National Science Center [N N301 707040 to Z.G., UMO-2013/08/A/ST5/00295, UMO-2011/03/B/NZ1/00576and UMO-2011/0/B/ST5/011098 to R.K.], Ministry of Science and Higher Education [N N301 255536 to Z.G.], Ministry of Science and Higher Education, European Fund for Regional Development [UDA-POIG.02.01.00–30–182/09 to Z.G.], EU COST action MP0802 [STSM n◦11227 to M.M.], and Fonds de la Recherche Scientifique-FNRS [research associate position and FRFC grant 2.4528.11 to V.G.]

Subjects
Circular dichroism, RNA Folding, Stereochemistry, Electrospray ionization, Biology, 010402 general chemistry, G-quadruplex, 01 natural sciences, 03 medical and health sciences, Trinucleotide Repeats, Structural Biology, Genetics, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Molecule, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Structural motif, Gene, 030304 developmental biology, 0303 health sciences, RNA, Molecular biology, 0104 chemical sciences, NMR spectra database, G-Quadruplexes, HeLa Cells
Abstract

Trinucleotide repeats are microsatellite sequences that are polymorphic in length. Their expansion in specific genes underlies a number of neurodegenerative disorders. Using ultraviolet-visible, circular dichroism, nuclear magnetic resonance (NMR) spectroscopies and electrospray ionization mass spectrometry, the structural preferences of RNA molecules composed of two and four repeats of AGG, CGG and UGG in the presence of K+, Na+ and NH4+ were analysed. (AGG)2A, (AGG)4A, p(UGG)2U and p(UGG)4U strongly prefer folding into G-quadruplexes, whereas CGG-containing sequences can adopt different types of structure depending on the cation and on the number of repeats. In particular, the two-repeat CGG sequence folds into a G-quadruplex in potassium buffer. We also found that each G-quadruplex fold is different: A:(G:G:G:G)A hexads were found for (AGG)2A, whereas mixed G:C:G:C tetrads and U-tetrads were observed in the NMR spectra of G(CGG)2C and p(UGG)2U, respectively. Finally, our NMR study highlights the influence of the strand sequence on the structure formed, and the influence of the intracellular environment on the folding. Importantly, we highlight that although potassium ions are prevalent in cells, the structures observed in the HeLa cell extract are not always the same as those prevailing in biophysical studies in the presence of K+ ions.

Published
2014
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5. Mitotic B-type cyclins are differentially regulated by phytohormones and during yellow lupine nodule development

Author

Joanna Jeleńska, Eva Kondorosi, Joanna Deckert, Andrzej B. Legocki, Institute of Bioorganic Chemistry [Poznań], Polska Akademia Nauk = Polish Academy of Sciences (PAN), Laboratory of Plant Ecophysiology, Adam Mickiewicz University in Poznań (UAM), Institut des sciences du végétal (ISV), and Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, Plant Science, Biology, 01 natural sciences, 03 medical and health sciences, chemistry.chemical_compound, Auxin, Botany, Gene expression, Genetics, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, Gene, Mitosis, 030304 developmental biology, Cyclin, chemistry.chemical_classification, 0303 health sciences, food and beverages, General Medicine, Meristem, Cell cycle, Cell biology, chemistry, Cytokinin, Agronomy and Crop Science, 010606 plant biology & botany
Abstract

The progression of cell cycle in eukaryotes is controlled by protein complexes composed of p34 protein kinase and cyclin subunits. Recently, we have described four B1 type mitotic cyclin in yellow lupine. The presence of several closely related cyclin genes within the same plant species raised the question about tissue specificity of respective cyclins or their different regulations by plant-specific signals. Therefore, we examined the expression pattern of four B1 cyclins in various lupine tissues, with special emphasis put on developing root nodules. We also studied the effect of phytohormones on the level of respective cyclin mRNAs. As expected, cyclin transcript accumulation was restricted to proliferating tissues. Detailed analysis by reverse transcription-PCR and using primers specific to each cyclin allow to establish that different genes are engaged in cell divisions of various meristematic tissues. All four genes were activated during nodule development, however, the Cyc3 and Cyc4 genes mostly at the early stages of nodulation, whereas the Cyc1 and Cyc2 genes within the mature nodule organs. The expression of cyclins was regulated differently by plant growth factors. Both auxin and cytokinin induced the Cyc1 and Cyc4 genes and their transcript level was also abundant within the root, shoot and floral meristems. The precise localisation of cyclin transcripts by in situ hybridisation revealed that lupine nodule meristem was active during the whole process of nodule development, even in old organs. © 2000 Elsevier Science Ireland Ltd. All rights reserved.

Published
2000
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6. Lupine leghemoglobin I: Expression in transgenic Lotus and tobacco tissues

Author

Wojciech M. Karlowski, Andrzej B. Legocki, Pawel M. Strozycki, Annik Petit, Yves Dessaux, Institute of Bioorganic Chemistry [Poznań], Polska Akademia Nauk = Polish Academy of Sciences (PAN), Institut des sciences du végétal (ISV), and Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, MESH: Sequence Homology, Amino Acid, Lotus, GUS reporter system, Genetically modified crops, MESH: Base Sequence, 01 natural sciences, Hemoglobins, Glucuronic Acid, MESH: Reverse Transcriptase Polymerase Chain Reaction, Lotus corniculatus, MESH: Tobacco, MESH: Phylogeny, Promoter Regions, Genetic, Phylogeny, 0303 health sciences, MESH: Leghemoglobin, Reverse Transcriptase Polymerase Chain Reaction, food and beverages, Plants, Genetically Modified, Leghemoglobin, MESH: Promoter Regions (Genetics), MESH: Hemoglobins, Biochemistry, MESH: Glucuronic Acid, MESH: Plants, Toxic, DNA, Complementary, Transgene, Molecular Sequence Data, MESH: Transformation, Genetic, Biology, MESH: Sequence Homology, Nucleic Acid, 03 medical and health sciences, Transformation, Genetic, Sequence Homology, Nucleic Acid, Tobacco, Genetics, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, Molecular Biology, Gene, 030304 developmental biology, MESH: Molecular Sequence Data, Base Sequence, Sequence Homology, Amino Acid, Promoter, MESH: DNA, Complementary, biology.organism_classification, Plants, Toxic, MESH: Plants, Genetically Modified, 010606 plant biology & botany
Abstract

The proximal parts of the promoters of the genes for symbiotic-type hemoglobins are generally conserved, but the promoter of the lbI gene of lupine (LulbI) shows some unusual structural features. It lacks typical organ-specific elements characteristic of all the leghemoglobin gene promoters described thus far. We have analysed its functional activity in transgenic Lotus corniculatus. A fusion construct between the lbI promoter and the GUS reporter gene was expressed mainly in the central zone of the root nodule, but the product was also detected in the non-nodule root zone and in roots in tissue culture. In roots of transgenic tobacco, the activity of the promoter was only 24% lower than in Lotus nodules. LulbI promoter activity was also detected in tobacco leaves. Lupine hemoglobin I has a higher sequence identity to symbiotic-type hemoglobins and thus it groups within the "Class II" hemoglobins.

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