C-terminal domain of nonhistone protein HMGB1 as a modulator of HMGB1–DNA structural interactions英文文献资料.docVIP

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C-terminal domain of nonhistone protein HMGB1 as a modulator of HMGB1–DNA structural interactions英文文献资料.doc

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C-terminal domain of nonhistone protein HMGB1 as a modulator of HMGB1–DNA structural interactions英文文献资料

Spectroscopy24(2010)361–366 DOI10.3233/SPE-2010-0457 IOSPress 361 C-terminaldomainofnonhistoneprotein HMGB1asamodulatorofHMGB1–DNA structuralinteractions andV.Vorob’ev E.Chikhirzhinaa,?,??,A.Polyanichkoa,b,?,Z.Leonenkoc,???,H.Wieserc a aInstituteofCytology,RussianAcademyofSciences,St.Petersburg,Russia bDepartmentofMolecularBiophysics,FacultyofPhysicsofSt.PetersburgStateUniversityand InstituteofCytologyRAS,St.Petersburg,Russia cDepartmentofChemistry,UniversityofCalgary,Calgary,Alberta,Canada Abstract.TheHMGB1protein(HighMobilityGroupprotein1)participatesintheformationoffunctionallysigni?cantDNA- proteincomplexes.HMGB1proteincontainstwoDNA-bindingdomainsandnegativelychargedC-terminalregion.Thelatter consistsofcontinuoussequenceofdicarboxylicaminoacidsresidues.StructuralchangesinDNA-proteincomplexeswerestud- iedbycirculardichroismspectroscopy(CD)andatomicforcemicroscopy(AFM).NaturalHMGB1andrecombinantprotein HMGB1(A+B)lackednegativelychargedC-terminalregionwereused.TheDNA–HMGB1(A+B)complexesdemonstrate anunusuallyhighopticalactivityin150mMNaClsolutions.AFMofthelattercomplexesshows,thatatthelowconcentra- tionofHMGB1inthecomplextheproteinisdistributedalongDNAinarandomway.IncreaseofHMGB1contentleadsto cooperativeinteractionandaredistributionoftheboundproteinmoleculesonDNAisobserved.Basedonthedataobtainedwe concludethatprotein–proteininteractionsplayakeyroleintheformationofhighlyorderedDNA–HMGB1complexes.Itwas shownthatC-terminaldomainmodulatetheinteractionsofDNAwithHMGB1protein.WesuggestthattheC-terminaldomain ofHMGB1alsomodulatesthe“packing”ofHMGB1moleculesontheDNA. Keywords:DNA,chromatin,HMGB1,circulardichroism,VCD 1. Introduction Structural-functional relationships in chromatin have been studied for manyyears. Amongnumer- ousDNA-proteincomplexesdeterminingchromatinfunctioning[4]themostintriguingonesinmany respectsarethosebetweenDNAandHMGB-domainproteins.HMGB-proteinsarethemembersofa largefamilyofHMG(HighMobility Group)non-histone chromatinpr