Purification and characterisation of the TnsB protein of Tn7: a transposition protein that binds to the ends of Tn7.
AUTOR(ES)
Tang, Y
RESUMO
Tn7, a large bacterial transposon encodes 5 proteins required for its transposition. We report a rapid and easy purification of one of these proteins, TnsB, from an overexpression strain. This protein was shown to bind to the ends of Tn7, in a bandshift assay, in two distinct stages as a function of protein concentration. DNasel footprinting at each end of Tn7 showed that the TnsB recognition sequence, a set of 22 bp repeats, plus Tn7 termini are protected. Binding of TnsB appeared cooperative but was only observed above a threshold concentration of protein. ATP and Mg2+ had no effect on the pattern of protection, nor did addition of other Tn7-encoded proteins. Hydroxyl radical footprinting, performed at the right end, showed that TnsB binds preferentially to one side of the DNA helix.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=328340Documentos Relacionados
- Interaction of the Tn7-encoded transposition protein TnsB with the ends of the transposon.
- Purification and characterization of TnsC, a Tn7 transposition protein that binds ATP and DNA.
- Identification of a transposon Tn7-dependent DNA-binding activity that recognizes the ends of Tn7.
- DNA sequence analysis of five genes; tnsA, B, C, D and E, required for Tn7 transposition.
- Tn7 recognizes transposition target structures associated with DNA replication using the DNA-binding protein TnsE