Rapid Analysis of Saccharomyces cerevisiae Genome Rearrangements by Multiplex Ligation–Dependent Probe Amplification 英文参考文献.docVIP

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Rapid Analysis of Saccharomyces cerevisiae Genome Rearrangements by Multiplex Ligation–Dependent Probe Amplification 英文参考文献.doc

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Rapid Analysis of Saccharomyces cerevisiae Genome Rearrangements by Multiplex Ligation–Dependent Probe Amplification 英文参考文献

RapidAnalysisofSaccharomycescerevisiaeGenome RearrangementsbyMultiplexLigation–Dependent ProbeAmplification JasonE.Chan1,2,3,RichardD.Kolodner2,3 * 1BioinformaticsandSystemsBiologyGraduateProgram,UniversityofCaliforniaSanDiego,LaJolla,California,UnitedStatesofAmerica,2LudwigInstituteforCancer Research,CancerCenterandDepartmentsofMedicineandCellularandMolecularMedicine,Moores–UCSDCancerCenter,SchoolofMedicine,UniversityofCaliforniaSan Diego,LaJolla,California,UnitedStatesofAmerica,3InstituteofGenomicMedicine,SchoolofMedicine,UniversityofCaliforniaSanDiego,LaJolla,California,United StatesofAmerica Abstract Aneuploidyandgrosschromosomalrearrangements(GCRs)canleadtogeneticdiseasesandthedevelopmentofcancer. We previouslydemonstrated thatintroduction of therepetitive retrotransposon Ty912onto anonessential chromosome armofSaccharomycescerevisiaeledtoincreasedgenomeinstabilitypredominantlyduetoincreasedratesofformationof monocentric nonreciprocal translocations. In this study, we adapted Multiplex Ligation–dependent Probe Amplification (MLPA)toanalyzealargenumbersoftheseGCRs.UsingMLPA,wefoundthatthedistributionoftranslocationsinducedby thepresenceofTy912inawild-typestrainwasnonrandomandthatthemajorityofthesetranslocationsweremediatedby onlysixtranslocationtargetsonfourdifferentchromosomes,eventhoughtherewere254potentialTy-relatedtranslocation targetsintheS.cerevisiaegenome.WhilethemajorityofTy912-mediatedtranslocationsresultedfromRAD52-dependent recombination, we observed a number of nonreciprocal translocations mediated by RAD52-independent recombination between Ty1 elements. The formation of these RAD52-independent translocations did not require the Rad51 or Rad59 homologous pairing proteins or the Rad1–Rad10 endonuclease complex that processes branched DNAs during recombination. Finally, we found that defects in ASF1-RTT109–dependent acetylation of histone H3 lysine residue 56 (H3K56)resultedinincreasedaccumulationofbothGCRsandwhole-chromosomeduplications,andresulted