EB1 Recognizes the Nucleotide State of Tubulin in the Microtubule Lattice 英文参考文献.docVIP

EB1RecognizestheNucleotideStateofTubulininthe MicrotubuleLattice MarijaZanic,JeffreyH.Stear¤,AnthonyA.Hyman,JonathonHoward* MaxPlanckInstituteofMolecularCellBiologyandGenetics,Dresden,Germany Abstract Plus-end-trackingproteins(+TIPs)arelocalizedatthefast-growing,orplusend,ofmicrotubules,andlinkmicrotubuleends to cellular structures. One of the best studied +TIPs is EB1, which forms comet-like structures at the tips of growing microtubules. The molecular mechanisms by which EB1 recognizes and tracks growing microtubule ends are largely unknown.However,oneclueisthatEB1canbinddirectlytoamicrotubuleendintheabsenceofotherproteins.Herewe use an in vitro assay for dynamic microtubule growth with two-color total-internal-reflection-fluorescence imaging to investigate binding of mammalian EB1 to both stabilized and dynamic microtubules. We find that under conditions of microtubule growth, EB1 not only tip tracks, as previously shown, but also preferentially recognizes the GMPCPP microtubulelatticeasopposedtotheGDPlattice.TheinteractionofEB1withtheGMPCPPmicrotubulelatticedependson the E-hook of tubulin, as well as the amount of salt in solution. The ability to distinguish different nucleotide states of tubulininmicrotubulelatticemaycontributetotheend-trackingmechanismofEB1. Citation: ZanicM, Stear JH, HymanAA,Howard J (2009) EB1 Recognizesthe Nucleotide Stateof Tubulinin the Microtubule Lattice.PLoS ONE 4(10):e7585. doi:10.1371/journal.pone.0007585 Editor:RobinCharlesMay,UniversityofBirmingham,UnitedKingdom ReceivedJune12,2009;AcceptedSeptember29,2009;PublishedOctober23,2009 Copyright: ? 2009 Zanic et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:MZissupportedbytheHumanFrontierinScienceProgram()Cross-DisciplinaryFellowship(LT00248/2008-C).Thefundershadnorolein studydesign,datacollectionandana