C. elegans BLOC-1 Functions in Trafficking to Lysosome-Related Gut Granules 英文参考文献.docVIP

C.elegansBLOC-1FunctionsinTraffickingtoLysosome- RelatedGutGranules GregJ.Hermann1,2*,EmilyScavarda1,AllisonM.Weis1,DanielS.Saxton1,LauraL.Thomas2, RebeccaSalesky1,HannahSomhegyi1,ThomasP.Curtin2,AlecBarrett2,OliviaK.Foster2,AnnaliseVine2, KatherineErlich2,ElizabethKwan2,BeverleyM.Rabbitts2,KailaWarren1 1DepartmentofBiology,LewisandClarkCollege,Portland,Oregon,UnitedStatesofAmerica,2PrograminBiochemistryandMolecularBiology,LewisandClarkCollege, Portland,Oregon,UnitedStatesofAmerica Abstract ThehumandiseaseHermansky-Pudlaksyndromeresultsfromdefectivebiogenesisoflysosome-relatedorganelles(LROs) andcanbecausedbymutationsinsubunitsoftheBLOC-1complex.HereweshowthatC.elegansglo-2andsnpn-1 ,despite relativelylowlevelsofaminoacididentity,encodePallidinandSnapinBLOC-1subunithomologues,respectively.BLOC-1 subunitinteractionsinvolvingPallidinandSnapinwereconservedforGLO-2andSNPN-1.Mutationsinglo-2andsnpn-1,or RNAitargeting5otherBLOC-1subunithomologuesinageneticbackgroundsensitizedforglo-2function,ledtodefectsin thebiogenesisoflysosome-relatedgutgranules.TheseresultsindicatethattheBLOC-1complexisconservedinC.elegans. ToaddressthefunctionofC.elegansBLOC-1,weassessedtheintracellularsortingofCDF-2::GFP,LMP-1,andPGP-2togut granules.WevalidatedtheirutilitybyanalyzingtheirmislocalizationinintestinalcellslackingthefunctionofAP-3,which participatesinanevolutionarilyconservedsortingpathwaytoLROs.BLOC-1(2)intestinalcellsmissortedgutgranulecargo to the plasma membrane and conventional lysosomes and did not have obviously altered function or morphology of organellescomposingtheconventionallysosomeproteinsortingpathway.DoublemutantanalysisandcomparisonofAP- 3(2)andBLOC-1(2)phenotypesrevealedthatBLOC-1hassomefunctionsindependentoftheAP-3adaptorcomplexin traffickingtogutgranules.WediscusssimilaritiesanddifferencesofBLOC-1activityinthebiogenesisofgutgranulesas comparedtomammalianmelanosomes,whereBLOC-1hasbeenmostextensivelystudiedforitsroleinsortingtoLROs.Our work opens up the opportunity to