Screen for ISG15-crossreactive Deubiquitinases 英文参考文献.docVIP

ScreenforISG15-crossreactiveDeubiquitinases Andre′ Catic1,2.¤,EddaFiebiger3.,GregoryA.Korbel2,Danie¨lBlom4,PaulJ.Galardy5*,HiddeL.Ploegh2* 1PrograminImmunology,HarvardMedicalSchool,Boston,Massachusetts,UnitedStatesofAmerica,2WhiteheadInstituteforBiomedicalResearch, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America, 3GI Cell Biology, Children’s Hospital, Boston, Massachusetts,UnitedStatesofAmerica,4Merck,Rahway,NewJersey,UnitedStatesofAmerica,5MayoClinic,PediatricandAdolescentMedicine, Rochester,Minnesota,UnitedStatesofAmerica Background. The family of ubiquitin-like molecules (UbLs) comprises several members, each of which has sequence, structural, or functional similarity to ubiquitin. ISG15 is a homolog of ubiquitin in vertebrates and is strongly upregulated followinginductionbytypeIinterferon.ISG15canbecovalentlyattachedtoproteins,analogoustoubiquitinationandwith actualsupportofubiquitinconjugatingfactors.SpecificproteasesareabletoreversemodificationwithubiquitinorUbLsby hydrolyzing the covalent bond between their C-termini and substrate proteins. The tail regions of ubiquitin and ISG15 are identical and we therefore hypothesized that promiscuous deubiquitinating proteases (DUBs) might exist, capable of recognizingbothubiquitinandISG15.Results.Wehaveclonedandexpressed22humanDUBs,representingthemajorclades oftheUSPproteasefamily.UtilizingsuicideinhibitorsbasedonubiquitinandISG15,wehaveidentifiedUSP2,USP5(IsoT1), USP13 (IsoT3), and USP14 as ISG15-reactive proteases, in addition to the bona fide ISG15-specific protease USP18 (UBP43). USP14isaproteasome-associatedDUB,anditsISG15isopeptidaseactivityincreaseswhencomplexedwiththeproteasome. Conclusions. By evolutionary standards, ISG15 is a newcomer among the UbLs and it apparently not only utilizes the conjugatingbutalsothedeconjugatingmachineryofitsmoreestablishedrelativeubiquitin.Functionaloverlapbetweenthese two posttranslational modifiers might therefore be more extensive t