Studies on a "jumping gene machine": higher-order nucleoprotein complexes in Mu DNA transposition.
Studies in my lab have focused on DNA transposition in the bacterial virus, Mu. In vitro studies have shown that Mu DNA transposition is a three-step process involving DNA breakage, strand transfer and DNA replication. In the first step, a nick is introduced at each end of the transposon. The liberated 3'-OH groups subsequently attack a target DNA molecule resulting in strand transfer. The transposon DNA, now covalently linked to the target, is finally replicated to generate the transposition end-product, referred to as a cointegrate. The DNA cleavage and strand transfer reactions are mediated by a "jumping gene machine" or transpososomes, which we discovered in 1987. They are assembled by bringing together three different DNA regions via a process involving multiple protein-DNA and protein-protein interactions. The action of four different proteins is required in addition to protein-induced DNA bending or wrapping to overcome the intrinsic stiffness of DNA, which would ordinarily prohibit the assembly of such a structure. Transpososome assembly is a gradual process involving multiple steps with an inherent flexibility whereby alternate pathways can be used in the assembly process, biasing the reaction towards completion under different conditions.
Chaconas
G
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Department of Biochemistry, University of Western Ontario, London, Canada. chaconas@julian.uwo.ca
D010583:Bacteriophage mu / Q000235:genetics* / Q000378:metabolism; D004261:DNA Replication; D004251:DNA Transposable Elements; D004742:Enhancer Elements, Genetic; D009698:Nucleoproteins / Q000378:metabolism*
D004251:DNA Transposable Elements; D009698:Nucleoproteins