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Insertion Sequences
Insertion Sequences or IS elements are the simplest form of mobile element in bacteria. They
are normal constituents of many bacterial chromosomes, bacterial plasmids and bacterial
virus genomes.
IS elements consist of a relatively short (700-1500 bp) DNA segment flanked by a 10-40 bp
inverted terminal repeat (ITR) sequence. A complete IS element codes for the protein
(transposase) that catalyses the transposition event. Thus, transposition requires that the IS
element carry a promoter recognized by the RNA polymerase of the host cell. Typically the
gene for the transposase is the only gene within the element.
Molecules of the transposase bind to the ITR sequences at the ends of the IS and bring the
two ends together into a complex, so that the IS element is looped out. Then it cuts both
DNA strands to excise the element. Subsequently, the DNA strands of the vacated site are
re-connected. Molecules of transposase remain attached to both ends of the excised IS
element. Next, the IS element with bound transposase makes staggered cuts in the two
strands of the new target site DNA and the IS element is inserted by ligating one strand of
the element to the target DNA at either end. Finally, the single strand gaps at either end
are filled (by hcellular enzymes) to create the characteristic Direct Terminal Repeat (DTR)
target site duplication that is characteristic of transposable elements.
The transposase is "trans-acting". This means that the transposase expressed from a
complete IS element in one location may mobilize transposition of a defective IS element
elsewhere in the cell which lacks a functional transposase gene; but only if the defective IS
element has an ITR sequence recognized by the transposase produced by the complete
element. The terms "autonomous" and "non-autonomous" are sometimes used to distinguish
complete and defective versions of IS elements.
Composite Transposons
These consist of two copies of the same IS element flanking variable amounts of other DNA
sequences coding for one or several genes with diverse functions, unrelated to transposition.
The best-known transposons carry antibiotic resistance genes.
The diagram below compares the typical structure of the IS50 element with the transposon
Tn5. Tn5 carries 3 antibiotic resistance genes flanked 2 copies of IS50.
IRL
transposase
IRR
16 bp INVERTED REPEAT!
(one mismatch)
Homologous Recombination
Multiple copies of the same transposable element in the same cell are substrates for
homologous recombination events that may lead to DNA deletions, sequence inversions, or
fusion of separate DNA molecules. For example, homologous recombination between copies of
the same IS element in a conjugal plasmid and the bacterial chromosome leads to formation
of Hfr strains, as shown below.
IS
IS
F+
IS
IS
Hfr