Principles of MicroRNA–Target Recognition 英文参考文献.docVIP

o Openaccess,freelyavailableonline PL S BIOLOGY PrinciplesofMicroRNA–TargetRecognition Julius Brennecke[,Alexander Stark[,Robert B.Russell, Stephen M. Cohen* EuropeanMolecularBiologyLaboratory,Heidelberg,Germany MicroRNAs(miRNAs)areshortnon-codingRNAsthatregulategeneexpressioninplantsandanimals.Althoughtheir biologicalimportancehasbecomeclear,howtheyrecognizeandregulatetargetgenesremainslesswellunderstood. Here, we systematically evaluate the minimal requirements for functional miRNA–target duplexes in vivo and distinguish classes of target sites with different functional properties. Target sites can be grouped into two broad categories.59dominantsiteshavesufficientcomplementaritytothemiRNA59endtofunctionwithlittleornosupport frompairingtothemiRNA39end.Indeed,siteswith39pairingbelowtherandomnoiselevelarefunctionalgivena strong59end.Incontrast,39compensatorysiteshaveinsufficient59pairingandrequirestrong39pairingforfunction. Wepresentexamplesandgenome-widestatisticalsupporttoshowthatbothclassesofsitesareusedinbiologically relevant genes. We provide evidence that an average miRNA has approximately 100 target sites, indicating that miRNAs regulate a large fraction of protein-coding genes and that miRNA 39 ends are key determinants of target specificitywithinmiRNAfamilies. Citation:BrenneckeJ,StarkA,RussellRB,CohenSM(2005)PrinciplesofmicroRNA–targetrecognition.PLoSBiol3(3):e85. number of predicted targets have proven to be functional whensubjectedtoexperimentaltests[19,26,27,29]. Introduction MicroRNAs (miRNAs) are small non-coding RNAs that serveaspost-transcriptionalregulatorsofgeneexpressionin plants and animals. They act by binding to complementary sites on target mRNAs to induce cleavage or repression of productivetranslation(reviewedin[1,2,3,4]).Theimportance of miRNAs for development is highlighted by the fact that theycompriseapproximately1%ofgenesinanimals,andare often highly conserved across a wide range of species (e.g., [5,6,7]). Further, mutations in p