Gene Conversion Transfers the GAF-A Domain of Phosphodiesterase TbrPDEB1 to One Allele of TbrPDEB2 of Trypanosoma brucei 英文参考文献.docVIP

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Gene Conversion Transfers the GAF-A Domain of Phosphodiesterase TbrPDEB1 to One Allele of TbrPDEB2 of Trypanosoma brucei 英文参考文献.doc

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Gene Conversion Transfers the GAF-A Domain of Phosphodiesterase TbrPDEB1 to One Allele of TbrPDEB2 of Trypanosoma brucei 英文参考文献

GeneConversionTransferstheGAF-ADomainof PhosphodiesteraseTbrPDEB1toOneAlleleofTbrPDEB2 ofTrypanosomabrucei StefanKunz,EdithLuginbuehl,ThomasSeebeck* InstituteofCellBiology,UniversityofBern,Bern,Switzerland Abstract Background:Chromosome9ofTrypanosomabruceicontainstwocloselyspaced,verysimilaropenreadingframesforcyclic nucleotide specific phosphodiesterases TbrPDEB1 and TbrPDEB2. They are separated by 2379bp, and both code for phosphodiesteraseswithtwoGAFdomainsintheirN-terminalmoietiesandacatalyticdomainattheC-terminus. MethodsandFindings:ThecurrentstudyrevealsthatintheLister427strainofT.brucei,thesetwogeneshaveundergone gene conversion, replacing the coding region for the GAF-A domain of TbrPDEB2 by the corresponding region of the upstreamgeneTbrPDEB1.Asaconsequence,thesestrainsexpresstwoslightlydifferentversionsofTbrPDEB2.TbrPDEB2a representsthewild-typephosphodiesterase,whileTbrPDEB2brepresentstheproductoftheconvertedgene.Earlierwork onthesubcellularlocalizationofTbrPDEB1andTbrPDEB2haddemonstratedthatTbrPDEB1ispredominantlylocatedinthe flagellum, whereas TbrPDEB2 partially locates tothe flagellum but largely remains in the cellbody. The currentfindings raisedthequestionofwhetherthisduallocalizationofTbrPDEB2mayreflectthetwoalleles.Toresolvethis,TbrPDEB2of strainSTIB247thatishomozygousforTbrPDEB2awastaggedinsitu,anditsintracellularlocalizationwasanalyzed. Conclusions: The results obtained were very similar to those found earlier with Lister427, indicating that the dual localizationofTbrPDEB2reflectsitstruefunctionandisnotsimplyduetothepresenceofthetwodifferentalleles.Notably, thegeneconversioneventisuniquefortheLister427strainandallitsderivatives.Basedonthisfinding,aconvenientPCR testhasbeendevelopedthatallowsthestringentdiscriminationbetweenLister-derivedstrainsthatarecommoninmany laboratoriesandotherisolates.Thetechniqueislikelyveryusefultoresolvequestionsaboutpotentialmix-upsofprecious fieldisolateswiththeubiquitousListerstrain. Citation:KunzS,LuginbuehlE,Seeb