Numt-Mediated Double-Strand Break Repair Mitigates Deletions during Primate Genome Evolution 英文参考文献.docVIP

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Numt-Mediated Double-Strand Break Repair Mitigates Deletions during Primate Genome Evolution 英文参考文献.doc

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Numt-Mediated Double-Strand Break Repair Mitigates Deletions during Primate Genome Evolution 英文参考文献

Numt-MediatedDouble-StrandBreakRepairMitigates DeletionsduringPrimateGenomeEvolution EinatHazkani-Covo1*,ShayCovo2 1NationalEvolutionarySynthesisCenter,Durham,NorthCarolina,UnitedStatesofAmerica,2LaboratoryofMolecularGenetics,ChromosomeStabilitySection,National InstituteofEnvironmentalHealthSciences,NationalInstitutesofHealth,ResearchTrianglePark,NorthCarolina,UnitedStatesofAmercia Abstract Non-homologous end joining (NHEJ) is the major mechanism of double-strand break repair (DSBR) in mammalian cells. NHEJhastraditionallybeeninferredfromexperimentalsystemsinvolvinginduceddoublestrandbreaks(DSBs).Whetheror not the spectrum of repair events observed in experimental NHEJ reflects the repair of natural breaks by NHEJ during chromosomal evolution is an unresolved issue. In primate phylogeny, nuclear DNA sequences of mitochondrial origin, numts,areinsertedintonaturallyoccurringchromosomalbreaksviaNHEJ.Thus,numtintegrationsitesharborevidencefor themechanismsthatactonthegenomeoverevolutionarytimescales.Wehaveidentified35and55lineage-specificnumts inthehumanandchimpanzeegenomes,respectively,usingtherhesusmonkeygenomeasanoutgroup.Onehundredand fiftytwonumt-chromosomefusionpointswereclassifiedbasedontheirrepairpatterns.Repairinvolvingmicrohomology andrepairleadingtonucleotideadditionsweredetected.Theserepairpatternsarewithintheexperimentallydetermined spectrumofclassicalNHEJ,suggestingthatinformationfromexperimentalsystemsisrepresentativeofbroadergeneticloci and end configurations. However, in incompatible DSBR events, small deletions always occur, whereas in 54% of numt integration events examined, no deletions were detected. Numts show a statistically significant reduction in deletion frequency,evenincomparisontoDSBRinvolvingfillerDNA.Therefore,numtsshowauniquemechanismofintegrationvia NHEJ.Sincethedeletionfrequencyduringnumtinsertionislow,nativeoverhangsofchromosomebreaksarepreserved, allowing us to determine that 24% of the analyzed breaks are cohesive with overhangs o