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Probing RNA structure by lead cleavage
1University of Chicago, Chicago, Illinois, USA.
Abstract:
Lead cleavage causes a transesterification reaction that breaks the 5',3'-phosphodiester backbone of RNA, leaving a 2',3'-cyclic phosphate and a 5'-hydroxyl. Since the efficiency of the reaction at the 2'-hydroxyl is related to steric and chemical constraints on particular 2'-hydroxyls embedded in the RNA, this reaction can be used to examine the structure of individual nucleotides within an RNA molecule. It is a sensitive probe of tertiary RNA structure, provided that Pb(2+)-binding sites are created in the tertiary structure.
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