Characteristics of MuA transposase-catalyzed processing of model transposon end DNA hairpin substrates.
Bacteriophage Mu uses non-replicative transposition for integration into the host's chromosome and replicative transposition for phage propagation. Biochemical and structural comparisons together with evolutionary considerations suggest that the Mu transposition machinery might share functional similarities with machineries of the systems that are known to employ a hairpin intermediate during the catalytic steps of transposition. Model transposon end DNA hairpin substrates were used in a minimal-component in vitro system to study their proficiency to promote Mu transpososome assembly and subsequent MuA-catalyzed chemical reactions leading to the strand transfer product. MuA indeed was able to assemble hairpin substrates into a catalytically competent transpososome, open the hairpin ends and accurately join the opened ends to the target DNA. The hairpin opening and transposon end cleavage reactions had identical metal ion preferences, indicating similar conformations within the catalytic center for these reactions. Hairpin length influenced transpososome assembly as well as catalysis: longer loops were more efficient in these respects. In general, MuA's proficiency to utilize different types of hairpin substrates indicates a certain degree of flexibility within the transposition machinery core. Overall, the results suggest that non-replicative and replicative transposition systems may structurally and evolutionarily be more closely linked than anticipated previously.
Saariaho
Anna-Helena
AH
Institute of Biotechnology, Program in Cellular Biotechnology. Viikki Biocenter, PO Box 56, Viikinkaari 9, 00014, University of Helsinki, Finland.
Savilahti
Harri
H
D010583:Bacteriophage mu / Q000201:enzymology*; D002384:Catalysis; D002413:Cations, Divalent; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004251:DNA Transposable Elements; D008670:Metals / Q000737:chemistry; D009690:Nucleic Acid Conformation; D013379:Substrate Specificity; D019895:Transposases / Q000378:metabolism*
D002413:Cations, Divalent; D004251:DNA Transposable Elements; D008670:Metals; D004247:DNA; D019895:Transposases