Functional analysis of a highly conserved abundant larval transcript-2 (alt-2) intron 2 repeat region of lymphatic filarial parasites.
Identifieur interne : 002A67 ( Main/Exploration ); précédent : 002A66; suivant : 002A68Functional analysis of a highly conserved abundant larval transcript-2 (alt-2) intron 2 repeat region of lymphatic filarial parasites.
Auteurs : Moorthy Sakthidevi [Inde] ; Sugeerappa Laxmanappa Hoti [Inde] ; Perumal Kaliraj [Inde]Source :
- Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases [ 1567-7257 ] ; 2014.
Descripteurs français
- KwdFr :
- ADN des helminthes (génétique), Animaux, Antigènes d'helminthe (biosynthèse), Antigènes d'helminthe (génétique), Antigènes d'helminthe (immunologie), Brugia malayi (génétique), Brugia malayi (immunologie), Cellules CHO, Cellules HepG2, Cellules Sf9, Cricetulus, Filariose lymphatique (parasitologie), Gènes rapporteurs (génétique), Humains, Introns (génétique), Larve (génétique), Liaison aux protéines, Lignée cellulaire, Luciferases (génétique), Protéines d'helminthes (génétique), Protéines nucléaires (métabolisme), Protéines recombinantes (biosynthèse), Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Régulation de l'expression des gènes, Répétitions minisatellites (génétique), Spodoptera, Structure tertiaire des protéines, Test de retard de migration électrophorétique, Wuchereria bancrofti (génétique), Wuchereria bancrofti (immunologie), Échappement immunitaire, Éléments silenceurs transcriptionnels (génétique).
- MESH :
- biosynthèse : Antigènes d'helminthe, Protéines recombinantes.
- génétique : ADN des helminthes, Antigènes d'helminthe, Brugia malayi, Gènes rapporteurs, Introns, Larve, Luciferases, Protéines d'helminthes, Protéines recombinantes, Répétitions minisatellites, Wuchereria bancrofti, Éléments silenceurs transcriptionnels.
- immunologie : Antigènes d'helminthe, Brugia malayi, Protéines recombinantes, Wuchereria bancrofti.
- métabolisme : Protéines nucléaires.
- parasitologie : Filariose lymphatique.
- Animaux, Cellules CHO, Cellules HepG2, Cellules Sf9, Cricetulus, Humains, Liaison aux protéines, Lignée cellulaire, Régulation de l'expression des gènes, Spodoptera, Structure tertiaire des protéines, Test de retard de migration électrophorétique, Échappement immunitaire.
English descriptors
- KwdEn :
- Animals, Antigens, Helminth (biosynthesis), Antigens, Helminth (genetics), Antigens, Helminth (immunology), Brugia malayi (genetics), Brugia malayi (immunology), CHO Cells, Cell Line, Cricetulus, DNA, Helminth (genetics), Electrophoretic Mobility Shift Assay, Elephantiasis, Filarial (parasitology), Gene Expression Regulation, Genes, Reporter (genetics), Helminth Proteins (genetics), Hep G2 Cells, Humans, Immune Evasion, Introns (genetics), Larva (genetics), Luciferases (genetics), Minisatellite Repeats (genetics), Nuclear Proteins (metabolism), Protein Binding, Protein Structure, Tertiary, Recombinant Proteins (biosynthesis), Recombinant Proteins (genetics), Recombinant Proteins (immunology), Sf9 Cells, Silencer Elements, Transcriptional (genetics), Spodoptera, Wuchereria bancrofti (genetics), Wuchereria bancrofti (immunology).
- MESH :
- chemical , biosynthesis : Antigens, Helminth, Recombinant Proteins.
- chemical , genetics : Antigens, Helminth, DNA, Helminth, Helminth Proteins, Luciferases, Recombinant Proteins.
- chemical , immunology : Antigens, Helminth, Recombinant Proteins.
- genetics : Brugia malayi, Genes, Reporter, Introns, Larva, Minisatellite Repeats, Silencer Elements, Transcriptional, Wuchereria bancrofti.
- immunology : Brugia malayi, Wuchereria bancrofti.
- chemical , metabolism : Nuclear Proteins.
- parasitology : Elephantiasis, Filarial.
- Animals, CHO Cells, Cell Line, Cricetulus, Electrophoretic Mobility Shift Assay, Gene Expression Regulation, Hep G2 Cells, Humans, Immune Evasion, Protein Binding, Protein Structure, Tertiary, Sf9 Cells, Spodoptera.
Abstract
The filarial-specific protein abundant larval transcript-2 (ALT-2) is expressed exclusively in the infective larval stage (L3) and is a crucial protein for establishing immunopathogenesis in human hosts. The alt-2 gene has a conserved minisatellite repeat (29 or 27bp) in intron 2 (IR2) whose significance within lymphatic filarial species is unknown. Here, we report the role of IR2 in the regulation of alt-2 gene expression using an in vitro model. Using electrophoretic mobility shift assays, we identified the presence of a putative nuclear protein binding region within IR2. Subsequent transient expression experiments in eukaryotic cell lines demonstrated that the IR2 downregulated the expression of a downstream luciferase reporter gene, which was further validated with RT-PCR. We therefore identify IR2 as a suppressor element that regulates L3 stage-specific expression of alt-2.
DOI: 10.1016/j.meegid.2014.03.017
PubMed: 24681262
Affiliations:
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(genetics)</term><term>Electrophoretic Mobility Shift Assay</term><term>Elephantiasis, Filarial (parasitology)</term><term>Gene Expression Regulation</term><term>Genes, Reporter (genetics)</term><term>Helminth Proteins (genetics)</term><term>Hep G2 Cells</term><term>Humans</term><term>Immune Evasion</term><term>Introns (genetics)</term><term>Larva (genetics)</term><term>Luciferases (genetics)</term><term>Minisatellite Repeats (genetics)</term><term>Nuclear Proteins (metabolism)</term><term>Protein Binding</term><term>Protein Structure, Tertiary</term><term>Recombinant Proteins (biosynthesis)</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (immunology)</term><term>Sf9 Cells</term><term>Silencer Elements, Transcriptional (genetics)</term><term>Spodoptera</term><term>Wuchereria bancrofti (genetics)</term><term>Wuchereria bancrofti (immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN des helminthes 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qualifier="immunology" xml:lang="en"><term>Brugia malayi</term><term>Wuchereria bancrofti</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Nuclear Proteins</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Protéines nucléaires</term></keywords><keywords scheme="MESH" qualifier="parasitologie" xml:lang="fr"><term>Filariose lymphatique</term></keywords><keywords scheme="MESH" qualifier="parasitology" xml:lang="en"><term>Elephantiasis, Filarial</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>CHO Cells</term><term>Cell Line</term><term>Cricetulus</term><term>Electrophoretic Mobility Shift Assay</term><term>Gene Expression Regulation</term><term>Hep G2 Cells</term><term>Humans</term><term>Immune Evasion</term><term>Protein Binding</term><term>Protein Structure, Tertiary</term><term>Sf9 Cells</term><term>Spodoptera</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules CHO</term><term>Cellules HepG2</term><term>Cellules Sf9</term><term>Cricetulus</term><term>Humains</term><term>Liaison aux protéines</term><term>Lignée cellulaire</term><term>Régulation de l'expression des gènes</term><term>Spodoptera</term><term>Structure tertiaire des protéines</term><term>Test de retard de migration électrophorétique</term><term>Échappement immunitaire</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The filarial-specific protein abundant larval transcript-2 (ALT-2) is expressed exclusively in the infective larval stage (L3) and is a crucial protein for establishing immunopathogenesis in human hosts. The alt-2 gene has a conserved minisatellite repeat (29 or 27bp) in intron 2 (IR2) whose significance within lymphatic filarial species is unknown. Here, we report the role of IR2 in the regulation of alt-2 gene expression using an in vitro model. Using electrophoretic mobility shift assays, we identified the presence of a putative nuclear protein binding region within IR2. Subsequent transient expression experiments in eukaryotic cell lines demonstrated that the IR2 downregulated the expression of a downstream luciferase reporter gene, which was further validated with RT-PCR. We therefore identify IR2 as a suppressor element that regulates L3 stage-specific expression of alt-2.</div></front></TEI><affiliations><list><country><li>Inde</li></country></list><tree><country name="Inde"><noRegion><name sortKey="Sakthidevi, Moorthy" sort="Sakthidevi, Moorthy" uniqKey="Sakthidevi M" first="Moorthy" last="Sakthidevi">Moorthy Sakthidevi</name></noRegion><name sortKey="Hoti, Sugeerappa Laxmanappa" sort="Hoti, Sugeerappa Laxmanappa" uniqKey="Hoti S" first="Sugeerappa Laxmanappa" last="Hoti">Sugeerappa Laxmanappa Hoti</name><name sortKey="Kaliraj, Perumal" sort="Kaliraj, Perumal" uniqKey="Kaliraj P" first="Perumal" last="Kaliraj">Perumal Kaliraj</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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