Summary information and primary citation
- PDB-id
-
2xo6;
DSSR-derived features in text and
JSON formats
- Class
- DNA binding protein-DNA
- Method
- X-ray (1.9 Å)
- Summary
- Deinococcus radiodurans isdra2 transposase y132f mutant
complexed with left end recognition and cleavage site
- Reference
-
Hickman AB, James JA, Barabas O, Pasternak C, Ton-Hoang
B, Chandler M, Sommer S, Dyda F (2010): "DNA
Recognition and the Precleavage State During
Single-Stranded DNA Transposition in D. Radiodurans."
Embo J., 29, 3840. doi:
10.1038/EMBOJ.2010.241.
- Abstract
- Bacterial insertion sequences (ISs) from the
IS200/IS605 family encode the smallest known DNA
transposases and mobilize through single-stranded DNA
transposition. Transposition by one particular family
member, ISDra2 from Deinococcus radiodurans, is
dramatically stimulated upon massive γ irradiation. We have
determined the crystal structures of four ISDra2
transposase/IS end complexes; combined with in vivo
activity assays and fluorescence anisotropy binding
measurements, these have revealed the molecular basis of
strand discrimination and transposase action. The
structures also show that previously established structural
rules of target site recognition that allow different
specific sequences to be targeted are only partially
conserved among family members. Furthermore, we have
captured a fully assembled active site including the
scissile phosphate bound by a divalent metal ion cofactor
(Cd²(+)) that supports DNA cleavage. Finally, the observed
active site rearrangements when the transposase binds a
metal ion in which it is inactive provide a clear rationale
for metal ion specificity.
