Noncanonical DNA Motifs as Transactivation Targets by Wild Type and Mutant p53 英文参考文献.docVIP
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Noncanonical DNA Motifs as Transactivation Targets by Wild Type and Mutant p53 英文参考文献
NoncanonicalDNAMotifsasTransactivationTargetsby
WildTypeandMutantp53
JenniferJ.Jordan1,2,DanielMenendez1,AlbertoInga1,3,MaherNourredine1,DouglasBell1,MichaelA.
Resnick1,2
*
1Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, NIH, Research Triangle Park, North Carolina, United States of America,
2Curriculum inGenetics andMolecular Biology, University ofNorth Carolina at ChapelHill, ChapelHill,North Carolina,United States ofAmerica, 3Unit ofMolecular
MutagenesisandDNARepair,NationalInstituteforCancerResearch,IST,Genoa,Italy
Abstract
Sequence-specific binding by the human p53 master regulator is critical to its tumor suppressor activity in response to
environmentalstresses.p53bindsasatetramertotwodecamerichalf-sitesseparatedby0–13nucleotides(nt),originally
definedbytheconsensusRRRCWWGYYY(n=0–13)RRRCWWGYYY.Tobetterunderstandtheroleofsequence,organization,
andlevelofp53ontransactivationattargetresponseelements(REs)bywildtype(WT)andmutantp53,wedeconstructed
thefunctionalp53canonicalconsensussequenceusingbuddingyeastandhumancellsystems.Contrarytoearlyreportson
binding in vitro, small increases in distance between decamer half-sites greatly reduces p53 transactivation, as
demonstrated for the natural TIGER RE. This was confirmed with human cell extracts using a newly developed, semi–in
vitromicrospherebindingassay.Theseresultscontrastwiththesynergisticincreaseintransactivationfromapairofweak,
full-siteREsintheMDM2promoterthatareseparatedbyanevolutionaryconserved17bpspacer.Surprisingly,therecanbe
substantialtransactivationatnoncanonicalK-(asingledecamer)andL-sites,someofwhichwereoriginallyclassifiedas
biologicallyrelevantcanonicalconsensussequencesincludingPIDDandApaf-1.p53familymembersp63andp73yielded
similar results. Efficient transactivation from noncanonical elements requires tetrameric p53, and the presence of the
carboxyterminal,non-specificDNAbindingdomainenhancedtransactivationfromnoncanonicalsequences.Ourfindings
demonstratetha
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