Diacylglycerol-Stimulated Endocytosis of Transferrin in Trypanosomatids Is Dependent on Tyrosine Kinase Activity 英文参考文献.docVIP

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Diacylglycerol-Stimulated Endocytosis of Transferrin in Trypanosomatids Is Dependent on Tyrosine Kinase Activity 英文参考文献.doc

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Diacylglycerol-Stimulated Endocytosis of Transferrin in Trypanosomatids Is Dependent on Tyrosine Kinase Activity 英文参考文献

Diacylglycerol-StimulatedEndocytosisofTransferrinin TrypanosomatidsIsDependentonTyrosineKinase Activity SandeshSubramanya¤,KojoMensa-Wilmot* DepartmentofCellularBiology,UniversityofGeorgia,Athens,Georgia,UnitedStatesofAmerica Abstract Small molecule regulation of cell function is an understudied area of trypanosomatid biology. In Trypanosoma brucei diacylglycerol(DAG)stimulatesendocytosisoftransferrin(Tf).However,itisnotknownwhetherothertrypanosomatidae respondsimilarlytothelipid.Further,thebiochemicalpathwaysinvolvedinDAGsignalingtotheendocyticsysteminT. bruceiareunknown,astheparasitegenomedoesnotencodecanonicalDAGreceptors(e.g.C1-domains).Weestablished thatDAGstimulatesendocytosisofTfinLeishmaniamajor,andweevaluatedpossibleeffectorenzymesinthepathwaywith multiple approaches. First, a heterologously expressed glycosylphosphatidylinositol phospholipase C (GPI-PLC ) activated endocytosisofTf300%inL.major.Second,exogenousphorbolesterandDAGspromotedTfendocytosisin L.major .In searchofpossibleeffectorsofDAGsignaling,wediscoveredanovelC1-likedomain(i.e.C1_5)intrypanosomatids,andwe identified protein Tyr kinases (PTKs) linked with C1_5 domains in T. brucei, T. cruzi, and L. major. Consequently, we hypothesizedthattrypanosomePTKsmightbeeffectorenzymesforDAGsignaling.GeneraluptakeofTfwasreducedby inhibitorsofeitherSer/ThrorTyrkinases.However,DAG-stimulatedendocytosisofTfwasblockedonlybyaninhibitorof PTKs,inbothT.bruceiandL.major.Weconcludethat(i)DAGactivatesTfendocytosisinL.major,andthat(ii)PTKsare effectors of DAG-stimulated endocytosis of Tf in trypanosomatids. DAG-stimulated endocytosis of Tf may be a T. brucei adaptationtocompeteeffectivelywithhostcellsforvertebrateTfinblood,sinceDAGdoesnotenhanceendocytosisofTf inhumancells. Citation: Subramanya S, Mensa-Wilmot K (2010) Diacylglycerol-Stimulated Endocytosis of Transferrin in Trypanosomatids Is Dependent on Tyrosine Kinase Activity.PLoSONE5(1):e8538.doi:10.1371/journal.pone.0008538 Editor:PingWang,ResearchIns