Effect of DNA Groove Binder Distamycin A upon Chromatin Structure 英文参考文献.docVIP

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Effect of DNA Groove Binder Distamycin A upon Chromatin Structure 英文参考文献.doc

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Effect of DNA Groove Binder Distamycin A upon Chromatin Structure 英文参考文献

EffectofDNAGrooveBinderDistamycinAupon ChromatinStructure ParijatMajumder,DipakDasgupta* BiophysicsDivision,SahaInstituteofNuclearPhysics,Kolkata,WestBengal,India Abstract Background:DistamycinAisaprototypeminorgroovebinder,whichbindstoB-formDNA,preferentiallyatA/Trichsites. ExtensiveworkinthepastfewdecadeshascharacterizedthebindingatthelevelofdoublestrandedDNA.However,effect ofthesameonphysiologicalDNA,i.e.DNAcomplexedinchromatin,hasnotbeenwellstudied.Hereweelucidatefroma structuralperspective,theinteractionofdistamycinwithsolublechromatin,isolatedfromSprague-Dawleyrat. Methodology/Principal Findings: Chromatin is a hierarchical assemblage of DNA and protein. Therefore, in order to characterizetheinteractionofthesamewithdistamycin,wehaveclassifiedthesystemintovariouslevels,accordingtothe requirements of the method adopted, and the information to be obtained. Isothermal titration calorimetry has been employedtocharacterizethebindingatthelevelsofchromatin,chromatosomeandchromosomalDNA.Thermodynamic parametersobtainedthereof,identifyenthalpyasthedrivingforcefortheassociation,withcomparablebindingaffinityand freeenergyforchromatinandchromosomalDNA.Reactionenthalpiesatdifferenttemperatureswereutilizedtoevaluate thechangeinspecificheatcapacity(DCp),which,inturn,indicatedapossiblebindingassociatedstructuralchange.Ligand induced structural alterations have been monitored by two complementary methods - dynamic light scattering, and transmission electron microscopy. They indicate compaction of chromatin. Using transmission electron microscopy, we havevisualizedtheeffectofdistamycinuponchromatinarchitectureatdi-andtrinucleosomelevels.Ourresultselucidate thesimultaneousinvolvementoflinkerbendingandinternucleosomalanglecontractionincompactionprocessinducedby distamycin. Conclusions/Significance: We summarize here, for the first time, the thermodynamic parameters for the interaction of distamycin with soluble chromatin, and elucidate its effect on chromatin architecture. The s