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2018 ; 46
(8
): 3864-3877
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gab.com Text
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English Wikipedia
Role for first zinc finger of WT1 in DNA sequence specificity: Denys-Drash
syndrome-associated WT1 mutant in ZF1 enhances affinity for a subset of WT1
binding sites
#MMPMID29294058
Wang D
; Horton JR
; Zheng Y
; Blumenthal RM
; Zhang X
; Cheng X
Nucleic Acids Res
2018[May]; 46
(8
): 3864-3877
PMID29294058
show ga
Wilms tumor protein (WT1) is a Cys2-His2 zinc-finger transcription factor vital
for embryonic development of the genitourinary system. The protein contains a
C-terminal DNA binding domain with four tandem zinc-fingers (ZF1-4). An
alternative splicing of Wt1 can add three additional amino acids-lysine (K),
threonine (T) and serine (S)-between ZF3 and ZF4. In the -KTS isoform, ZF2-4
determine the sequence-specificity of DNA binding, whereas the function of ZF1
remains elusive. Three X-ray structures are described here for wild-type -KTS
isoform ZF1-4 in complex with its cognate DNA sequence. We observed four unique
ZF1 conformations. First, like ZF2-4, ZF1 can be positioned continuously in the
DNA major groove forming a 'near-cognate' complex. Second, while ZF2-4 make
base-specific interactions with one DNA molecule, ZF1 can interact with a second
DNA molecule (or, presumably, two regions of the same DNA molecule). Third, ZF1
can intercalate at the joint of two tail-to-head DNA molecules. If such
intercalation occurs on a continuous DNA molecule, it would kink the DNA at the
ZF1 binding site. Fourth, two ZF1 units can dimerize. Furthermore, we examined a
Denys-Drash syndrome-associated ZF1 mutation (methionine at position 342 is
replaced by arginine). This mutation enhances WT1 affinity for a guanine base.
X-ray crystallography of the mutant in complex with its preferred sequence
revealed the interactions responsible for this affinity change. These results
provide insight into the mechanisms of action of WT1, and clarify the fact that
ZF1 plays a role in determining sequence specificity of this critical
transcription factor.