TY - JOUR
T1 - Structure of the AvrBs3-DNA complex provides new insights into the initial thymine-recognition mechanism
AU - Stella, Stefano
AU - Molina, Rafael
AU - Yefimenko, Igor
AU - Prieto, Jesús
AU - Silva, George
AU - Bertonati, Claudia
AU - Juillerat, Alexandre
AU - Duchateau, Phillippe
AU - Montoya, Guillermo
PY - 2013/9/1
Y1 - 2013/9/1
N2 - Transcription activator-like effectors contain a DNA-binding domain organized in tandem repeats. The repeats include two adjacent residues known as the repeat variable di-residue, which recognize a single base pair, establishing a direct code between the dipeptides and the target DNA. This feature suggests this scaffold as an excellent candidate to generate new protein-DNA specificities for biotechnological applications. Here, the crystal structure of AvrBs3 (residues 152-895, molecular mass 82kDa) in complex with its target DNA sequence is presented, revealing a new mode of interaction with the initial thymine of the target sequence, together with an analysis of both the binding specificity and the thermodynamic properties of AvrBs3. This study quantifies the affinity and the specificity between AvrBs3 and its target DNA. Moreover, in vitro and in vivo analyses reveal that AvrBs3 does not show a strict nucleotide-binding preference for the nucleotide at the zero position of the DNA, widening the number of possible sequences that could be targeted by this scaffold.
AB - Transcription activator-like effectors contain a DNA-binding domain organized in tandem repeats. The repeats include two adjacent residues known as the repeat variable di-residue, which recognize a single base pair, establishing a direct code between the dipeptides and the target DNA. This feature suggests this scaffold as an excellent candidate to generate new protein-DNA specificities for biotechnological applications. Here, the crystal structure of AvrBs3 (residues 152-895, molecular mass 82kDa) in complex with its target DNA sequence is presented, revealing a new mode of interaction with the initial thymine of the target sequence, together with an analysis of both the binding specificity and the thermodynamic properties of AvrBs3. This study quantifies the affinity and the specificity between AvrBs3 and its target DNA. Moreover, in vitro and in vivo analyses reveal that AvrBs3 does not show a strict nucleotide-binding preference for the nucleotide at the zero position of the DNA, widening the number of possible sequences that could be targeted by this scaffold.
KW - gene targeting
KW - genetics
KW - protein-DNA interaction
UR - http://www.scopus.com/inward/record.url?scp=84883588736&partnerID=8YFLogxK
U2 - 10.1107/s0907444913016429
DO - 10.1107/s0907444913016429
M3 - Journal article
C2 - 23999294
AN - SCOPUS:84883588736
SN - 2059-7983
VL - 69
SP - 1707
EP - 1716
JO - Acta Crystallographica Section D: Structural Biology
JF - Acta Crystallographica Section D: Structural Biology
IS - 9
ER -