"Using a lacZ reporter gene inserted into a YAC construct, we demonstrate that WT1 is expressed in the early proepicardium, the epicardium and the subepicardial mesenchymal cells (SEMC)."

"Knock-out mice were shown to have defects in the urogenital system and to die at embryonic day E13.5, probably due to heart failure."

"Analysis of the observed hypoplastic kidneys demonstrate a continuous requirement for WT1 during nephrogenesis, in particular, in the formation of mature glomeruli."

"Finally, we show that the development of adrenal glands is also severely affected in partially rescued embryos."

"we demonstrate that reduced expression levels of WT1 result in either crescentic glomerulonephritis or mesangial sclerosis depending on the gene dosage."

"Strikingly, the two podocyte-specific genes nphs1 and podocalyxin are dramatically downregulated in mice with decreased levels of Wt1, suggesting that these two genes act downstream of Wt1."

"WT1 bound to several sites in this region of the promoter, as demonstrated by gel-shift analysis and DNase I footprinting, and functioned as a powerful repressor of PDGF-A transcription in vivo."

"WT1 and BASP1 associate within the nuclei of cells that naturally express both proteins."

"BASP1 can confer WT1 cosuppressor activity in transfection assays, and elimination of endogenous BASP1 expression augments transcriptional activation by WT1."

"Here we show that WT1 interacts with the novel splicing regulator RBM4."

"We found that overexpressed WT1(+KTS) abrogated this effect of RBM4 on splice-site selection, whereas WT1(-KTS) did not. We conclude that the (+KTS) form of WT1 is able to inhibit the effect of RBM4 on alternative splicing."

"Each protein was found to colocalize in nuclear speckles and to cosediment with supraspliceosomes in glycerol gradients."

"We demonstrate that depletion of WT1 by short-interfering RNAs leads to reduced DNMT3A in Wilms' tumour cells and human embryonal kidney-derived cell lines."

"Chromatin immunoprecipitation assays demonstrate WT1 recruitment to the DNMT3A promoter region and reporter assays confirm that WT1 directly transactivates DNMT3A expression."

"Using genome-wide promoter methylation arrays, we show that human embryonal kidney cells over-expressing WT1 acquire DNA methylation changes at specific gene promoters where DNMT3A recruitment is increased, with hypermethylation being associated with silencing of gene expression."

"Consequently, WT1 binds the same consensus sequence as Egr1/Zif268 (Stoll et al. 2007)"

"Egr1/Zif268 and WT1 showed slightly higher affinity for the fully methylated sequence by factors of ∼2.8 (Egr1) and ∼1.8 (WT1)"

"Both displayed high affinity for the sequence when C or 5 mC was present and much reduced affinity when 5 hmC or 5 fC was present"

"We found that WT1 can repress transcription of both the bcl-2 and c-myc promoters. This suggests that WT1 regulates bcl-2 and c-myc during renal development, and the loss of functional WT1 results in deregulation of bcl-2 and c-myc, contributing to tumor formation."

"Binding of WT1 to three high affinity sites in the 5' untranslated Pax-2 leader sequence was demonstrated by DNAseI footprinting analysis."

"Furthermore, co-transfection assays using CAT reporter constructs under the control of Pax-2 regulatory sequences demonstrated WT1-dependent transcriptional repression."

"By immunoprecipitation and protein microsequencing analysis, we have identified a major cellular protein associated with endogenous WT1 to be the inducible chaperone Hsp70."

"These observations indicate that Hsp70 is an important cofactor for the function of WT1, and suggest a potential role for this chaperone during kidney differentiation."

"Hsp70 is recruited to the characteristic subnuclear clusters that contain WT1."

"SRY (the sex-determining region of the Y chromosome) promoter was strongly activated by the WT1 isoforms without the KTS tripeptide, WT1(-)KTS, but was not activated by the WT1 isoforms with the KTS tripeptide, WT1(+)KTS, in all cells tested."

"The MüAdullerian-inhibiting substance (MIS) promoter was strongly repressed by WT1(-)KTS isoforms and more weakly repressed by the WT1(+)KTS isoforms in Sertoli cells but not in HeLa cells."

"The second alternative splicing site, which inserts the tripeptide KTS, alters the DNA binding capability."

"Domain architecture and nucleic-acid interactions: WT1 contains four C2H2 zinc fingers (exons 7-10) mediating high-affinity binding to GC-rich motifs such as 5'-GCG(T/G)GGGCG-3' in target promoters/enhancers"

"The -KTS isoform binds DNA sequence-specifically and drives transcriptional programs, whereas the +KTS isoform shows low DNA affinity, associates with spliceosomal components, and has RNA-binding/processing roles"

"WT1(+KTS) co-localizes with splicing speckles, interacts with splicing factors (U2AF65, WTAP), forms complexes with ZNF255 and BASP1 that participate in RNA maturation"

"Nephrogenesis and podocyte maintenance: WT1 binds and regulates kidney lineage and podocyte genes and is essential for nephron development and adult podocyte integrity"

"WT1 mRNA is overexpressed in >80% of AML; standardized ELN qRT-PCR thresholds are widely used for MRD risk stratification and post-transplant surveillance"

"The second, third, and fourth zinc finger domains insert deep into the major groove of DNA where they make base-specific contacts. Zinc fingers 2 and 3 are primarily responsible for most of the binding affinity and can function as a minimal DNA binding domain, while zinc finger 4 is indispensable for target specificity"

"Quantitative binding studies have demonstrated that recombinant WT1 zinc fingers bind to the dodecamer DNA sequence GCG-TGG-GCG-TGT with high affinity, exhibiting a dissociation constant (Kd) of 1.14 x 10^-9 M under physiological conditions. Thermodynamic analysis revealed that WT1-DNA interaction is an entropy-driven process"

"The molecular basis for the functional differences between +KTS and -KTS isoforms was elucidated by NMR structural studies demonstrating that the KTS insertion increases the flexibility of the linker between fingers 3 and 4, abrogating the binding of the fourth zinc finger to its cognate site in the DNA major groove"

"The physiological importance of maintaining the correct ratio of +KTS to -KTS isoforms (normally approximately 2:1) is dramatically illustrated by Frasier syndrome, in which mutations disrupting the +KTS splice site alter this ratio to 1:2"

"In the absence of p53, WT1 functions as a potent transcriptional activator of EGR1-like promoter sequences rather than as a repressor. WT1 expression can stabilize p53 protein, enhance its DNA binding to target sequences"

"Mechanistically, myristoylated BASP1 binds to nuclear phosphatidylinositol 4,5-bisphosphate (PIP2), which leads to the recruitment of PIP2 to the promoter regions of WT1-dependent target genes. This association with PIP2 is required for the interaction of BASP1 with histone deacetylase 1 (HDAC1), enabling chromatin deacetylation and gene silencing"

"WT1 binds to a specific responsive element in the nephrin promoter and activates transcription more than 10-fold. Transgenic mouse analysis confirmed that this WT1 binding site is required for podocyte-specific nephrin expression in vivo"

"The -KTS isoform of WT1, but not the +KTS form, directly binds to and transactivates the Sf1 promoter through four identified binding sites. WT1 also directly regulates the sex-determining gene SRY by activating its expression. Mutant WT1 proteins found in Denys-Drash syndrome patients fail to activate the SRY promoter"

"WT1 is expressed in the proepicardium and developing epicardium, where it is required for epithelial-to-mesenchymal transition (EMT) that generates cardiovascular progenitor cells. WT1 regulates epicardial EMT through multiple signaling pathways including canonical Wnt/beta-catenin signaling and retinoic acid signaling through direct regulation of Raldh2 expression"

"While zinc fingers 2, 3, and 4 are required for high-affinity DNA binding, zinc finger 1—which is dispensable for DNA binding—is more important for RNA binding. Consequently, the KTS insertion has minimal effect on RNA affinity"