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dc.contributor.authorHaley, Benjamin
dc.contributor.authorZamore, Phillip D.
dc.date2022-08-11T08:08:58.000
dc.date.accessioned2022-08-23T16:14:11Z
dc.date.available2022-08-23T16:14:11Z
dc.date.issued2004-06-01
dc.date.submitted2008-09-25
dc.identifier.citationNat Struct Mol Biol. 2004 Jul;11(7):599-606. Epub 2004 May 30. <a href="http://dx.doi.org/10.1038/nsmb780">Link to article on publisher's site</a>
dc.identifier.issn1545-9993 (Print)
dc.identifier.doi10.1038/nsmb780
dc.identifier.pmid15170178
dc.identifier.urihttp://hdl.handle.net/20.500.14038/33804
dc.description.abstractThe siRNA-directed ribonucleoprotein complex, RISC, catalyzes target RNA cleavage in the RNA interference pathway. Here, we show that siRNA-programmed RISC is a classical Michaelis-Menten enzyme in the presence of ATP. In the absence of ATP, the rate of multiple rounds of catalysis is limited by release of the cleaved products from the enzyme. Kinetic analysis suggests that different regions of the siRNA play distinct roles in the cycle of target recognition, cleavage, and product release. Bases near the siRNA 5' end disproportionately contribute to target RNA-binding energy, whereas base pairs formed by the central and 3' regions of the siRNA provide a helical geometry required for catalysis. Finally, the position of the scissile phosphate on the target RNA seems to be determined during RISC assembly, before the siRNA encounters its RNA target.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15170178&dopt=Abstract ">Link to article in PubMed</a>
dc.relation.urlhttp://dx.doi.org/10.1038/nsmb780
dc.subjectAdenosine Triphosphate; Animals; Base Sequence; Catalysis; DNA Primers; Drosophila melanogaster; Kinetics; Protein Binding; *RNA Interference; RNA, Small Interfering; Sequence Homology, Nucleic Acid
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleKinetic analysis of the RNAi enzyme complex
dc.typeJournal Article
dc.source.journaltitleNature structural and molecular biology
dc.source.volume11
dc.source.issue7
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/gsbs_sp/465
dc.identifier.contextkey638233
html.description.abstract<p>The siRNA-directed ribonucleoprotein complex, RISC, catalyzes target RNA cleavage in the RNA interference pathway. Here, we show that siRNA-programmed RISC is a classical Michaelis-Menten enzyme in the presence of ATP. In the absence of ATP, the rate of multiple rounds of catalysis is limited by release of the cleaved products from the enzyme. Kinetic analysis suggests that different regions of the siRNA play distinct roles in the cycle of target recognition, cleavage, and product release. Bases near the siRNA 5' end disproportionately contribute to target RNA-binding energy, whereas base pairs formed by the central and 3' regions of the siRNA provide a helical geometry required for catalysis. Finally, the position of the scissile phosphate on the target RNA seems to be determined during RISC assembly, before the siRNA encounters its RNA target.</p>
dc.identifier.submissionpathgsbs_sp/465
dc.contributor.departmentDepartment of Biochemistry and Molecular Pharmacology
dc.source.pages599-606


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