Conformational isomerization in phage Mu transpososome assembly: effects of the transpositional enhancer and of MuB.
Initiation of phage Mu DNA transposition requires assembly of higher order protein-DNA complexes called Mu transpososomes containing the two Mu DNA ends and MuA transposase tetramer. Mu transpososome assembly is highly regulated and involves multiple DNA sites for transposase binding, including a transpositional enhancer called the internal activation sequence (IAS). In addition, a number of protein cofactors participate, including the target DNA activator MuB ATPase. We investigated the impact of the assembly cofactors on the kinetics of transpososome assembly with the aim of deciphering the reaction steps that are influenced by the cofactors. The transpositional enhancer IAS appears to have little impact on the initial pairing of the two Mu end segments bound by MuA. Instead, it accelerates the post-synaptic conformational step(s) that converts the reversible complex to the stable transpososome. The transpososome assembly stimulation by MuB does not require its stable DNA binding activity, which appears critical for directing transposition to sites distant from the donor transposon.
Mizuuchi
M
M
Laboratory of Molecular Biology, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Mizuuchi
K
K
D010583:Bacteriophage mu / Q000737:chemistry* / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D004251:DNA Transposable Elements / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D005033:Ethylmaleimide / Q000494:pharmacology; D007536:Isomerism; D007700:Kinetics; D008954:Models, Biological; D011485:Protein Binding; D011487:Protein Conformation; D013997:Time Factors; D019895:Transposases / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism*
D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; C069215:MuB protein, Enterobacteria phage Mu; D014764:Viral Proteins; D004247:DNA; C114697:mu transposase; D019895:Transposases; D005033:Ethylmaleimide