PMID:12093750
| Citation |
Skelding, Z, Sarnovsky, R and Craig, NL (2002) Formation of a nucleoprotein complex containing Tn7 and its target DNA regulates transposition initiation. EMBO J. 21:3494-504 |
|---|---|
| Abstract |
Tn7 insertion into its specific target site, attTn7, is mediated by the proteins TnsA, TnsB, TnsC and TnsD. The double-strand breaks that separate Tn7 from the donor DNA require the Tns proteins, the transposon and an attTn7 target DNA, suggesting that a prerequisite for transposition is the formation of a nucleoprotein complex containing TnsABC+D, and these DNAs. Here, we identify a TnsABC+D transposon-attTn7 complex, and demonstrate that it is a transposition intermediate. We demonstrate that an interaction between TnsB, the transposase subunit that binds to the transposon ends, and TnsC, the target DNA-binding protein that controls the activity of the transposase, is essential for assembly of the TnsABC+D transposon-attTn7 complex. We also show that certain TnsB residues are required for recombination because they mediate a TnsB-TnsC interaction critical to formation of the TnsABC+D transposon-attTn7 complex. We demonstrate that TnsA, the other transposase subunit, which also interacts with TnsC, greatly stabilizes the TnsABC+D transposon-attTn7 complex. Thus multiple interactions between the transposase subunits, TnsA and TnsB, and the target-binding transposase activator, TnsC, control Tn7 transposition. |
| Links |
PubMed PMC126096 Online version:10.1093/emboj/cdf347 |
| Keywords |
Amino Acid Sequence; Bacterial Proteins/chemistry; Bacterial Proteins/physiology; Bacteriophage mu/physiology; DNA Transposable Elements/physiology; DNA, Bacterial/chemistry; DNA, Bacterial/metabolism; DNA-Binding Proteins/chemistry; DNA-Binding Proteins/physiology; Escherichia coli/genetics; Escherichia coli Proteins; Macromolecular Substances; Magnesium/physiology; Models, Genetic; Molecular Sequence Data; Nucleoproteins/chemistry; Protein Interaction Mapping; Protein Structure, Tertiary; Recombination, Genetic; Sequence Alignment; Sequence Homology, Amino Acid |
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