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abstracts: poster presentations

peter walter, president jonathan weissman, program chair

san francisco, california dec 3-7

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POSTERPRESENTATIONSTUESDAY

P2342
BoardNumber:B1316
TowardsUnderstandingtheCrystalStructureofWildTypePhenylalanineHydroxylase:TheHO
CountDiscrepancy.
O.Grosser1,J.Caradonna2;
1
Biotechnology,MassachusettsBayCommunityCollege,Wellesley,MA,2Chemistry,BostonUniversity,
Boston,MA

Phenylketonuria(PKU)isandautosomalrecessivediseasethataffects1/15000peopleintheUnited
States.ItarisesfromamutationinthePhenylalanineHydroxylase(PheH)genethatreducesitsactivity.
PheHisanenzymethatisfoundintheliver;withthehelpofthecoenzymepterin,itconvertsL
phenylalanineintoLtyrosine.HighamountsofLphenylalanineinthebloodistoxic;itcausesthe
degradationoftheneuronmyelinsheathswhichleadstomentalretardationandpermanentlylowers
onesIQ.
ThecrystalgraphicdataofwildtypePheHisnotconsistentwiththespectroscopicdata.Thecrystal
graphicdatasuggestsinmultipleactivesitegeometriesthatcontaindifferentamountsofHO.Weaim
tobringthesediscrepanciestolightbycountingtheamountofwatermoleculesinthewildtypeandthe
truncatedsequences.WealsoaimtodiscernthenumberofwatermoleculesintwoPheHmutants
(E286AandE330A).

Embryogenesis2

P2343
BoardNumber:B1318
ImplicationofImportin8inmousebraindevelopment.
G.NGANOU1,B.COUMANS1,T.GRISAR1,L.DENIJS2,B.LAKAYE1;
1
GIGANeurosciences,UniversityofLiege,Lige,Belgium,2MHeNsSchoolforMentalHealthand
Neuroscience,MaastrichtUniversity,Maastricht,Netherlands

Rationale:Regulationofnucleocytoplasmictransportofproteinsbythekaryopherinsuperfamilyis
criticalforcellphysiologyasitcontrolsmainyfundamentalprocessessuchasdivision,differenciation,
migrationandadaptationtoexternalenvironment.Besidethisfundamentalrole,the1and2
membersofthissuperfamilyarealsoimplicatedinmitosisandciliaryentryrespectively.Importin8
(IPO8),amemberofthekaryopherinfamily,isreportedtocontrolthetransportAgo2,cJunand
Smad4forexample,threeproteinsimportantforbraindevelopment.
Methods:ImmunostainingofHEKandhTERTcellsexpressingEGFPIPO8wereperformedwithtubulin
andacetylatedtubulinrespectively.InSituHybridization(ISH)atvariousembryonic(E12,E14,E18)
andpostnatalages(P5,P60)wereperformedusingdiglabeledriboprobes.InUteroElectroporation
(IUE)ofplasmidsexpressinganshRNAagainstmIPO8and/orahIPO8wereperformedatE14and
neuroblastmigrationwasanalyzedthreedayslaterbyimmunocytochemistryusinganantiEGFP
antibody.
Results:NocolocalisationofIPO8witheitherthemitoticspindle(inHEKcells)ortheprimarycilia(in
hTERTcells)couldbeobserved.SoitseemsIPO8onlyplaysaroleinnucleartransportofproteins.Then
wehaveassessedtheexpressionofIPO8inmousebrain.UsingISH,aclearexpressionofmIPO8mRNA
wasobservedinbrainswhatevertheagesalsowithdifferentlevels.AtE14,themRNAisexpressedin
thesubventricular/ventricularzone(SVZ/VZ),thecorticalplate(CP)andtheganglioniceminences.The
roleofIPO8oncerebralcortexdevelopment,andmorespecificallyradialmigration,wasinvestigatedby