An Experimental and Computational Study of the Effect of ActA Polarity on the Speed of Listeria monocytogenes Actin-based Motility 英文参考文献.docVIP

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An Experimental and Computational Study of the Effect of ActA Polarity on the Speed of Listeria monocytogenes Actin-based Motility 英文参考文献.doc

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An Experimental and Computational Study of the Effect of ActA Polarity on the Speed of Listeria monocytogenes Actin-based Motility 英文参考文献

AnExperimentalandComputationalStudyoftheEffect ofActAPolarityontheSpeedofListeriamonocytogenes Actin-basedMotility SusanneM.Rafelski¤.,JonathanB.Alberts*.,GarrettM.Odell CenterforCellDynamics,UniversityofWashington,FridayHarbor,Washington,UnitedStatesofAmerica Abstract Listeriamonocytogenesisapathogenicbacteriumthatmoveswithininfectedcellsandspreadsdirectlybetweencellsby harnessingthecell’sdendriticactinmachinery.Thismotilityisdependentonexpressionofasinglebacterialsurfaceprotein, ActA,aconstitutivelyactiveArp2,3activator,andhasbeenwidelystudiedasabiochemicalandbiophysicalmodelsystem foractin-basedmotility.Dendriticactinnetworkdynamicsareimportantforcellprocessesincludingeukaryoticcellmotility, cytokinesis,andendocytosis.HereweexperimentallyalteredthedegreeofActApolarityonapopulationofbacteriaand madeuseofanActA-RFPfusiontodeterminetherelationshipbetweenActAdistributionandspeedofbacterialmotion.We found a positive linear relationship for both ActA intensity and polarity with speed. We explored the underlying mechanismsofthisdependencewithtwodistinctlydifferentquantitativemodels:adetailedagent-basedmodelinwhich each actin filament and branched network is explicitly simulated, and a three-state continuum model that describes a simplified relationship between bacterial speed and barbed-end actin populations. In silico bacterial motility required a cooperativerestrainingmechanismtoreconstituteourobservedspeed-polarityrelationship,suggestingthatkineticfriction betweenactinfilamentsandthebacterialsurface,arestrainingforcepreviouslyneglectedinmotilitymodels,isimportantin determining the effect of ActA polarity on bacterial motility. The continuum model was less restrictive, requiring only a filament number-dependent restraining mechanism to reproduce our experimental observations. However, seemingly rationalassumptionsinthecontinuummodel,e.g.anaveragepropulsiveforceperfilament,wereinvalidatedbyfurther analysiswiththeagent-basedmodel.Wefoundthattheaveragecontributiontomot