Criss-crossed interactions between the enhancer and the att sites of phage Mu during DNA transposition.
A bipartite enhancer sequence (composed of the O1 and O2 operator sites) is essential for assembly of the functional tetramer of phage Mu transposase (MuA) on supercoiled DNA substrates. A three-site interaction (LER) between the left (L) and right (R) ends of Mu (att sites) and the enhancer (E) precedes tetramer assembly. We have dissected the role of the enhancer in tetramer assembly by using two transposase proteins that have a common att site specificity, but are distinct in their enhancer specificity. The activity of these proteins on substrates containing hybrid enhancers reveals a 'criss-crossed' pattern of interaction between att and enhancer sites. The left operator, O1, of the enhancer interacts specifically with the transposase subunit at the R1 site (within the right att sequence) that is responsible for cleaving the left end of Mu. The right operator, O2, shows a preferential interaction with the transposase subunit at the L1 site (within the left att sequence) that is responsible for cleaving the right end of Mu.
Jiang
H
H
Department of Microbiology and Institute of Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712, USA.
Yang
J Y
JY
Harshey
R M
RM
D001287:Attachment Sites, Microbiological / Q000235:genetics / Q000502:physiology*; D010583:Bacteriophage mu / Q000201:enzymology / Q000235:genetics*; D001665:Binding Sites; D004274:DNA, Recombinant / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics / Q000502:physiology*; D009154:Mutation; D009875:Operator Regions, Genetic / Q000502:physiology; D010957:Plasmids / Q000235:genetics; D011533:Proviruses / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D013379:Substrate Specificity; D019895:Transposases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C114697:mu transposase; D019895:Transposases