PSSA-2, a Membrane-Spanning Phosphoprotein of Trypanosoma brucei, Is Required for Efficient Maturation of Infection 英文参考文献.docVIP

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PSSA-2, a Membrane-Spanning Phosphoprotein of Trypanosoma brucei, Is Required for Efficient Maturation of Infection 英文参考文献.doc

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PSSA-2, a Membrane-Spanning Phosphoprotein of Trypanosoma brucei, Is Required for Efficient Maturation of Infection 英文参考文献

PSSA-2,aMembrane-SpanningPhosphoproteinof Trypanosomabrucei,IsRequiredforEfficientMaturation ofInfection CristinaM.Fragoso1¤,GabrielaSchumannBurkard1,MichaelOberle1,2,ChristinaKunzRenggli2 ,Karen Hilzinger1,IsabelRoditi1* 1Institutfu¨rZellbiologie,Universita¨t Bern,Bern,Switzerland,2SwissTropicalInstitute,Basel,Switzerland Abstract The coat of Trypanosoma brucei consists mainly of glycosylphosphatidylinositol-anchored proteins that are present in severalmillioncopiesandarecharacteristicofdefinedstagesofthelifecycle.Whilethesemajorcomponentsofthecoatsof bloodstream forms and procyclic (insect midgut) forms are well characterised, very little is known about less abundant stage-regulated surface proteins and their roles in infection and transmission. By creating epitope-tagged versions of procyclic-specificsurfaceantigen2(PSSA-2)wedemonstratedthatitisamembrane-spanningproteinthatisexpressedby severaldifferentlifecyclestagesintsetseflies,butnotbyparasitesinthemammalianbloodstream.Incommonwithother membrane-spanning proteins in T. brucei, PSSA-2 requires its cytoplasmic domain in order to exit the endoplasmic reticulum.CorrectlocalisationofPSSA-2requiresphosphorylationofacytoplasmicthreonineresidue(T305),amodification that depends on the presence of TbMAPK4. Mutation of T305 to alanine (T305A) has no effect on the localisation of the proteinincellsthatexpresswildtypePSSA-2.Incontrast,thisproteinislargelyintracellularwhenexpressedinanullmutant background.AvariantwithaT305Dmutationgivesstrongsurfaceexpressioninboththewildtypeandnullmutant,but slowsgrowthofthecells,suggestingthatitmayfunctionasadominantnegativemutant.ThePSSA-2nullmutantexhibits noperceptiblephenotypeincultureandisfullycompetentatestablishingmidgutinfectionsintsetse,butisdefectivein colonising the salivary glands and the production of infectious metacyclic forms. Given the protein’s structure and the effectsofmutationofT305onproliferationandlocalisation,wepostulatethatPSSA-2mightsenseandtransmitsignalsth