DNA recognition sites activate MuA transposase to perform transposition of non-Mu DNA.
Mu transposition occurs within a large protein-DNA complex called a transpososome. This stable complex includes four subunits of MuA transposase, each contacting a 22-base pair recognition site located near an end of the transposon DNA. These MuA recognition sites are critical for assembling the transpososome. Here we report that when concentrations of Mu DNA are limited, the MuA recognition sites permit assembly of transpososomes in which non-Mu DNA substitutes for some of the Mu sequences. These "hybrid" transpososomes are stable to competitor DNA, actively transpose the non-Mu DNA, and produce transposition products that had been previously observed but not explained. The strongest activator of non-Mu transposition is a DNA fragment containing two MuA recognition sites and no cleavage site, but a shorter fragment with just one recognition site is sufficient. Based on our results, we propose that MuA recognition sites drive assembly of functional transpososomes in two complementary ways. Multiple recognition sites help physically position MuA subunits in the transpososome plus each individual site allosterically activates transposase.
Goldhaber-Gordon
Ilana
I
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Williams
Tanya L
TL
Baker
Tania A
TA
D000495:Allosteric Site; D010584:Bacteriophage phi X 174 / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D004247:DNA / Q000378:metabolism*; D004251:DNA Transposable Elements; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D004789:Enzyme Activation; D008954:Models, Biological; D008969:Molecular Sequence Data; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D019895:Transposases / Q000378:metabolism*
D004251:DNA Transposable Elements; D004247:DNA; C114697:mu transposase; D019895:Transposases