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PDBsum entry 2kvg
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Transcription
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PDB id
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2kvg
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References listed in PDB file
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Key reference
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Title
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Structure and DNA binding characteristics of the three-Cys(2)his(2) domain of mouse testis zinc finger protein.
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Authors
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C.C.Chou,
Y.C.Lou,
T.K.Tang,
C.Chen.
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Ref.
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Proteins, 2010,
78,
2202-2212.
[DOI no: ]
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PubMed id
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Abstract
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The C-terminal three-Cys(2)His(2) zinc-finger domain (TZD) of mouse testis
zinc-finger protein binds to the 5'-TGTACAGTGT-3' at the Aie1 (aurora-C)
promoter with high specificity. Interestingly, the primary sequence of TZD is
unique, possessing two distinct linkers, TGEKP and GAAP, and distinct residues
at presumed DNA binding sites at each finger, especially finger 3. A K(d) value
of approximately 10(-8) M was obtained from surface plasmon resonance analysis
for the TZD-DNA complex. NMR structure of the free TZD showed that each zinc
finger forms a typical beta beta alpha fold. On binding to DNA, chemical shift
perturbations and the R(2) transverse relaxation rate in finger 3 are
significantly smaller than those in fingers 1 and 2, which indicates that the
DNA binding affinity in finger 3 is weaker. Furthermore, the shift perturbations
between TZD in complex with the cognate DNA and its serial mutants revealed that
both ADE7 and CYT8, underlined in 5'-ATATGTACAGTGTTAT-3', are critical in
specific binding, and the DNA binding in finger 3 is sequence independent.
Remarkably, the shift perturbations in finger 3 on the linker mutation of TZD
(GAAP mutated to TGEKP) were barely detected, which further indicates that
finger 3 does not play a critical role in DNA sequence-specific recognition. The
complex model showed that residues important for DNA binding are mainly located
on positions -1, 2, 3, and 6 of alpha-helices in fingers 1 and 2. The DNA
sequence and nonsequence-specific bindings occurring simultaneously in TZD
provide valuable information for better understanding of protein-DNA recognition.
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