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Comprehensive analysis of the KSHV MiRNA targetome by Ago-HITS-CLIP
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Title: Comprehensive analysis of the KSHV MiRNA targetome by Ago-HITS-CLIP
Series Title: Infectious Agents and Cancer
Physical Description: Book
Language: English
Creator: Haecker, Irina
Gay, Lauren
Morse, Alison
McCrory, Marty
Yang, Yajie
Hu, Jianhong
McIntyre, Lauren
Renne, Rolf
Publisher: BioMed Central
Publication Date: 2012
 Notes
General Note: From 13th International Conference on Malignancies in AIDS and Other Acquired Immunodeficiencies (ICMAOI) Bethesda, MD, USA. 7-8 November 2011
General Note: Oral presentation
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Source Institution: University of Florida
Holding Location: University of Florida
Rights Management: All rights reserved by the source institution.
Resource Identifier: doi - 10.1186/1750-9378-7-S1-O4
System ID: AA00012409:00001

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ORALPRESENTATION OpenAccessComprehensiveanalysisoftheKSHVMiRNA targetomebyAgo-HITS-CLIPIrinaHaecker,LaurenGay,AlisonMorse,MartyMcCrory,YajieYang,JianhongHu,LaurenMcIntyre,RolfRenne*From 13thInternationalConferenceonMalignanciesinAIDSandOtherAcquiredImmunodeficiencies (ICMAOI) Bethesda,MD,USA.7-8November2011Thegamma-herpesvirusKaposi sSarcoma-associated Herpesvirus(KSHV)istheetiologicalagentofKaposi s Sarcoma(KS),PrimaryEffusionLymphoma(PEL)anda subsetofMulticentricCastleman sDisease(MCD)in immunocompromisedindivid uals.Asallherpesviruses KSHVhasalatentandalyticlifecycle.Malignantcellsin KS,PEL,andMCDarelatentlyinfectedwithKSHV.Interestingly,thevirusencodes12miRNAgenesgivingriseto 25maturemiRNAsthatarepredominantlyexpressedduringlatency,i.e.inmalignantcells.This,togetherwiththe increasingevidencefortheinvolvementofmiRNAsin cancer,suggestsapotentialrolefortheKSHVmiRNAsin viraltumorigenesis.However,todate,verylittleisknown aboutthefunctionoftheseviralmiRNAs.Toaddressthe questionweperformedAgo-HITS-CLIP[1]usingthe anti-Agoantibody2A8[2]toisolateRISCcomplexesfrom KSHV-infectedlymphomac ells(BCBL1,BC3).RNAs extractedfromthesecomplexeswereanalyzedbyIllumina sequencingtoidentifyviralandcellularmiRNAsandtheir targetgenes.Thesearchforcanonicalseedsequence matches(nt2-7)oftheKSHVmiRNAswithinthe mRNA-derivedsequencingtagsrevealedmorethan1000 cellulartargets.Geneontologyanalysisrevealedthat KSHVmiRNAtargetsareenrichedingenesinvolvedin apoptosis,lymphocyteactivation,cellcycleregulation,and transcriptionalcontrol.Importantly,wereproducibly obtainedclustersofreadsonexperimentallyconfirmed KSHVmiRNAtargetsitesinseveralknowntargetgenes (e.g.Bach1,BCLAF1,andTHBS1).Newtargetgenesare intheprocessofexperimentalvalidation,with4confirmedtargetssofar:TP53INP1,TPD52,ANXA2, C/EBPB.Targetanalysisfornon-canonicalseedsaswellas forcellularmiRNAsiscurrentlyongoing.Published:19April2012 References1.ChiSW,ZangJB,MeleA,DarnellRB: ArgonauteHITS-CLIPdecodes microRNA-mRNAinteractionmaps. Nature 2009, 460(7254) :479-86. 2.NelsonPT,DePlanell-SaguerM,LamprinakiS,KiriakidouM,ZhangP, O DohertyU,MourelatosZ: Anovelmonoclonalantibodyagainsthuman ArgonauteproteinsrevealsunexpectedcharacteristicsofmiRNAsin humanbloodcells. RNA 2007, 13(10) :1787-92.doi:10.1186/1750-9378-7-S1-O4 Citethisarticleas: Haecker etal .: ComprehensiveanalysisoftheKSHV MiRNAtargetomebyAgo-HITS-CLIP. InfectiousAgentsandCancer 2012 7 (Suppl1):O4. Submit your next manuscript to BioMed Central and take full advantage of: Convenient online submission Thorough peer review No space constraints or color gure charges Immediate publication on acceptance Inclusion in PubMed, CAS, Scopus and Google Scholar Research which is freely available for redistribution Submit your manuscript at www.biomedcentral.com/submit *Correspondence:rrenne@ufl.edu DepartmentofMolecularGeneticsandMicrobiology,UniversityofFlorida, Gainesville,FL,USAHaecker etal InfectiousAgentsandCancer 2012, 7 (Suppl1):O4 http://www.infectagentscancer.com/content/7/S1/O4 2012Haeckeretal;licenseeBioMedCentralLtd.ThisisanopenaccessarticledistributedunderthetermsoftheCreativeCommons AttributionLicense(http://creativecommons.org/licenses/by/2.0),whichpermitsunrestricteduse,distribution,andreproductionin anymedium,providedtheoriginalworkisproperlycited.


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supplement Proceedings of the 13th International Conference on Malignancies in AIDS and Other Acquired Immunodeficiencies (ICMAOI)editor Geraldina Domingueznote Meeting abstractsconference 13th International Conference on Malignancies in AIDS and Other Acquired Immunodeficiencies (ICMAOI)location Bethesda, MD, USAdate-range 7-8 November 2011url http://www.capconcorp.com/meeting/2011/13thICMAOI/index.aspissn 1750-9378
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cpyrt 2012collab Haecker et al; licensee BioMed Central Ltd.This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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The gamma-herpesvirus Kaposi’s Sarcoma-associated Herpesvirus (KSHV) is the etiological agent of Kaposi’s Sarcoma (KS), Primary Effusion Lymphoma (PEL) and a subset of Multicentric Castleman’s Disease (MCD) in immunocompromised individuals. As all herpesviruses KSHV has a latent and a lytic life cycle. Malignant cells in KS, PEL, and MCD are latently infected with KSHV. Interestingly, the virus encodes 12 miRNA genes giving rise to 25 mature miRNAs that are predominantly expressed during latency, i.e. in malignant cells. This, together with the increasing evidence for the involvement of miRNAs in cancer, suggests a potential role for the KSHV miRNAs in viral tumorigenesis. However, to date, very little is known about the function of these viral miRNAs. To address the question we performed Ago-HITS-CLIP abbrgrp
abbr bid B1 1
using the anti-Ago antibody 2A8
B2 2
to isolate RISC complexes from KSHV-infected lymphoma cells (BCBL1, BC3). RNAs extracted from these complexes were analyzed by Illumina sequencing to identify viral and cellular miRNAs and their target genes. The search for canonical seed sequence matches (nt 2-7) of the KSHV miRNAs within the mRNA-derived sequencing tags revealed more than 1000 cellular targets. Gene ontology analysis revealed that KSHV miRNA targets are enriched in genes involved in apoptosis, lymphocyte activation, cell cycle regulation, and transcriptional control. Importantly, we reproducibly obtained clusters of reads on experimentally confirmed KSHV miRNA target sites in several known target genes (e.g. Bach1, BCLAF1, and THBS1). New target genes are in the process of experimental validation, with 4 confirmed targets so far: TP53INP1, TPD52, ANXA2, C/EBPB. Target analysis for non-canonical seeds as well as for cellular miRNAs is currently ongoing.
bm
refgrp Argonaute HITS-CLIP decodes microRNA-mRNA interaction mapsChiSWZangJBMeleADarnellRBNature20094607254479lpage 86pubidlist pmcid 2733940pmpid link fulltext 19536157A novel monoclonal antibody against human Argonaute proteins reveals unexpected characteristics of miRNAs in human blood cellsNelsonPTDe Planell-SaguerMLamprinakiSKiriakidouMZhangPO’DohertyUMourelatosZRNA2007131017879210.1261/rna.646007198680517720879