Functional analysis of a highly conserved abundant larval transcript-2 (alt-2) intron 2 repeat region of lymphatic filarial parasites.
Identifieur interne : 001540 ( PubMed/Checkpoint ); précédent : 001539; suivant : 001541Functional 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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pubmed:24681262Le document en format XML
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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 (génétique)</term><term>Animaux</term><term>Antigènes d'helminthe (biosynthèse)</term><term>Antigènes d'helminthe (génétique)</term><term>Antigènes d'helminthe (immunologie)</term><term>Brugia malayi (génétique)</term><term>Brugia malayi (immunologie)</term><term>Cellules CHO</term><term>Cellules HepG2</term><term>Cellules Sf9</term><term>Cricetulus</term><term>Filariose lymphatique (parasitologie)</term><term>Gènes rapporteurs (génétique)</term><term>Humains</term><term>Introns (génétique)</term><term>Larve (génétique)</term><term>Liaison aux protéines</term><term>Lignée cellulaire</term><term>Luciferases (génétique)</term><term>Protéines d'helminthes (génétique)</term><term>Protéines nucléaires (métabolisme)</term><term>Protéines recombinantes (biosynthèse)</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (immunologie)</term><term>Régulation de l'expression des gènes</term><term>Répétitions minisatellites (génétique)</term><term>Spodoptera</term><term>Structure tertiaire des protéines</term><term>Test de retard de migration électrophorétique</term><term>Wuchereria bancrofti (génétique)</term><term>Wuchereria bancrofti (immunologie)</term><term>Échappement immunitaire</term><term>Éléments silenceurs transcriptionnels (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Antigens, Helminth</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antigens, Helminth</term><term>DNA, Helminth</term><term>Helminth Proteins</term><term>Luciferases</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antigens, Helminth</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Antigènes d'helminthe</term><term>Protéines 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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><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">24681262</PMID><DateCreated><Year>2014</Year><Month>05</Month><Day>09</Day></DateCreated><DateCompleted><Year>2015</Year><Month>01</Month><Day>12</Day></DateCompleted><DateRevised><Year>2014</Year><Month>05</Month><Day>09</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1567-7257</ISSN><JournalIssue CitedMedium="Internet"><Volume>24</Volume><PubDate><Year>2014</Year><Month>Jun</Month></PubDate></JournalIssue><Title>Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases</Title><ISOAbbreviation>Infect. Genet. Evol.</ISOAbbreviation></Journal><ArticleTitle>Functional analysis of a highly conserved abundant larval transcript-2 (alt-2) intron 2 repeat region of lymphatic filarial parasites.</ArticleTitle><Pagination><MedlinePgn>177-82</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.meegid.2014.03.017</ELocationID><ELocationID EIdType="pii" ValidYN="Y">S1567-1348(14)00099-9</ELocationID><Abstract><AbstractText>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.</AbstractText><CopyrightInformation>Copyright © 2014 Elsevier B.V. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Sakthidevi</LastName><ForeName>Moorthy</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Centre for Biotechnology, Anna University, Chennai, Tamil Nadu 600 025, India.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hoti</LastName><ForeName>Sugeerappa Laxmanappa</ForeName><Initials>SL</Initials><AffiliationInfo><Affiliation>Vector Control Research Centre, Puducherry 605 006, India.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kaliraj</LastName><ForeName>Perumal</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>Centre for Biotechnology, Anna University, Chennai, Tamil Nadu 600 025, India. Electronic address: pk.kaliraj@gmail.com.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2014</Year><Month>03</Month><Day>26</Day></ArticleDate></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>Infect Genet Evol</MedlineTA><NlmUniqueID>101084138</NlmUniqueID><ISSNLinking>1567-1348</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C521555">ALT-2 protein, Brugia malayi</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000947">Antigens, Helminth</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018509">DNA, Helminth</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D015801">Helminth Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D009687">Nuclear Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.13.12.-</RegistryNumber><NameOfSubstance UI="D008156">Luciferases</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000947" MajorTopicYN="N">Antigens, Helminth</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D017178" MajorTopicYN="N">Brugia malayi</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D016466" MajorTopicYN="N">CHO Cells</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002460" MajorTopicYN="N">Cell Line</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D003412" MajorTopicYN="N">Cricetulus</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018509" MajorTopicYN="N">DNA, Helminth</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D024202" MajorTopicYN="N">Electrophoretic Mobility Shift Assay</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004605" MajorTopicYN="N">Elephantiasis, Filarial</DescriptorName><QualifierName UI="Q000469" MajorTopicYN="Y">parasitology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005786" MajorTopicYN="N">Gene Expression Regulation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017930" MajorTopicYN="N">Genes, Reporter</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015801" MajorTopicYN="N">Helminth Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D056945" MajorTopicYN="N">Hep G2 Cells</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D057131" MajorTopicYN="N">Immune Evasion</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007438" MajorTopicYN="N">Introns</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D007814" MajorTopicYN="N">Larva</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008156" MajorTopicYN="N">Luciferases</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018598" MajorTopicYN="N">Minisatellite Repeats</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D009687" MajorTopicYN="N">Nuclear Proteins</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011485" MajorTopicYN="N">Protein Binding</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017434" MajorTopicYN="N">Protein Structure, Tertiary</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D061987" MajorTopicYN="N">Sf9 Cells</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D037941" MajorTopicYN="N">Silencer Elements, Transcriptional</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018411" MajorTopicYN="N">Spodoptera</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014958" MajorTopicYN="N">Wuchereria bancrofti</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Evolution</Keyword><Keyword MajorTopicYN="N">Gene regulation</Keyword><Keyword MajorTopicYN="N">Intron DNA repeat</Keyword><Keyword MajorTopicYN="N">Silencer</Keyword><Keyword MajorTopicYN="N">Transcription</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2013</Year><Month>05</Month><Day>09</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2014</Year><Month>03</Month><Day>12</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2014</Year><Month>03</Month><Day>17</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2014</Year><Month>4</Month><Day>1</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2014</Year><Month>4</Month><Day>1</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2015</Year><Month>1</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">24681262</ArticleId><ArticleId IdType="pii">S1567-1348(14)00099-9</ArticleId><ArticleId IdType="doi">10.1016/j.meegid.2014.03.017</ArticleId></ArticleIdList></PubmedData></pubmed><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 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