title: Nicking by transesterification: the reaction catalysed by a relaxase.
abstract: DNA relaxases play an essential role in the initiation and termination of conjugative DNA transfer. Purification and characterization of relaxases from several plasmids has revealed the reaction mechanism: relaxases nick duplex DNA in a site- and strand-specific manner by catalysing a transesterification. The product of the reaction is a nicked double-stranded DNA molecule with a sequestered 3'-OH and the relaxase covalently bound to the 5' end of the cleaved strand via a phosphotyrosyl linkage. The relaxase-catalysed transesterification is isoenergetic and reversible; a second transesterification ligates the nicked DNA. However, the covalent nucleoprotein complex is relatively long-lived, a property that is likely to be essential for its role as an intermediate in the process of conjugative DNA transfer. Subsequent unwinding of the nicked DNA intermediate is required to produce the single strand of DNA transferred to the recipient cell. This reaction is catalysed by a DNA helicase, an activity intrinsic to the relaxase protein in some, but not all, plasmid systems. The first relaxase-catalysed transesterification is essential for initiation of conjugative strand transfer, whereas the second is presumably required for termination of the process. The relaxase, in conjunction with several auxiliary proteins, forms the relaxation complex or relaxosome first described nearly 30 years ago as being associated with conjugative and mobilizable plasmids.
authors.affiliation: Department of Biology, University of North Carolina at Chapel Hill, 27599, USA.
authors.forename: D R
authors.initials: DR
authors.lastname: Byrd
authors.forename: S W
authors.initials: SW
authors.lastname: Matson
mesh_terms: D000595:Amino Acid Sequence; D002384:Catalysis; D003227:Conjugation, Genetic; D004247:DNA / Q000378:metabolism*; D004254:DNA Nucleotidyltransferases; D004951:Esterification; D008969:Molecular Sequence Data; D010957:Plasmids
chemical_list: D004247:DNA; D004254:DNA Nucleotidyltransferases; C110340:DNA relaxase