Summary information and primary citation
- PDB-id
-
3zlj;
DSSR-derived features in text and
JSON formats
- Class
- DNA binding protein-DNA
- Method
- X-ray (3.1 Å)
- Summary
- Crystal structure of full-length e.coli DNA mismatch
repair protein muts d835r mutant in complex with gt
mismatched DNA
- Reference
-
Groothuizen FS, Fish A, Petoukhov MV, Reumer A, Manelyte
L, Winterwerp HHK, Marinus MG, Lebbink JHG, Svergun DI,
Friedhoff P, Sixma TK (2013): "Using
Stable Muts Dimers and Tetramers to Quantitatively
Analyze DNA Mismatch Recognition and Sliding Clamp
Formation." Nucleic Acids Res.,
41, 8166. doi: 10.1093/NAR/GKT582.
- Abstract
- The process of DNA mismatch repair is initiated when
MutS recognizes mismatched DNA bases and starts the repair
cascade. The Escherichia coli MutS protein exists in an
equilibrium between dimers and tetramers, which has
compromised biophysical analysis. To uncouple these states,
we have generated stable dimers and tetramers,
respectively. These proteins allowed kinetic analysis of
DNA recognition and structural analysis of the full-length
protein by X-ray crystallography and small angle X-ray
scattering. Our structural data reveal that the
tetramerization domains are flexible with respect to the
body of the protein, resulting in mostly extended
structures. Tetrameric MutS has a slow dissociation from
DNA, which can be due to occasional bending over and
binding DNA in its two binding sites. In contrast, the
dimer dissociation is faster, primarily dependent on a
combination of the type of mismatch and the flanking
sequence. In the presence of ATP, we could distinguish two
kinetic groups: DNA sequences where MutS forms sliding
clamps and those where sliding clamps are not formed
efficiently. Interestingly, this inability to undergo a
conformational change rather than mismatch affinity is
correlated with mismatch repair.
