ABSTRACT
WT1, a critical regulator of kidney development, is a tumor suppressor for nephroblastoma but in some contexts functions as an oncogene. A limited number of direct transcriptional targets of WT1 have been identified to explain its complex roles in tumorigenesis and organogenesis. In this study we performed genome-wide screening for direct WT1 targets, using a combination of ChIP-ChIP and expression arrays. Promoter regions bound by WT1 were highly G-rich and resembled the sites for a number of other widely expressed transcription factors such as SP1, MAZ, and ZNF219. Genes directly regulated by WT1 were implicated in MAPK signaling, axon guidance, and Wnt pathways. Among directly bound and regulated genes by WT1, nine were identified in the Wnt signaling pathway, suggesting that WT1 modulates a subset of Wnt components and responsive genes by direct binding. To prove the biological importance of the interplay between WT1 and Wnt signaling, we showed that WT1 blocked the ability of Wnt8 to induce a secondary body axis during Xenopus embryonic development. WT1 inhibited TCF-mediated transcription activated by Wnt ligand, wild type and mutant, stabilized beta-catenin by preventing TCF4 loading onto a promoter. This was neither due to direct binding of WT1 to the TCF binding site nor to interaction between WT1 and TCF4, but by competition of WT1 and TCF4 for CBP. WT1 interference with Wnt signaling represents an important mode of its action relevant to the suppression of tumor growth and guidance of development.
Subject(s)
Genetic Testing , Genome/genetics , Signal Transduction/genetics , WT1 Proteins/metabolism , Wnt Proteins/metabolism , Animals , Base Sequence , Binding Sites , CREB-Binding Protein/metabolism , Cell Line, Tumor , Chromatin Immunoprecipitation , DNA/metabolism , Embryo, Nonmammalian/metabolism , Embryonic Development , Gene Expression Profiling , Gene Expression Regulation , Molecular Sequence Data , Oligonucleotide Array Sequence Analysis , Promoter Regions, Genetic/genetics , Protein Binding , TCF Transcription Factors/metabolism , Transcription, Genetic , Xenopus/embryologyABSTRACT
The radiographs of 37 incontinent adult male dogs with urethral sphincter mechanism incompetence were compared with those of 28 control dogs to determine if, as in the bitch, differences in bladder neck position and urethral length were implicated in the pathophysiology of urethral sphincter mechanism incompetence. Bladder neck position was significantly different; compared with continent dogs, incontinent animals were significantly more likely (P < 0.005) to have intrapelvic than intra-abdominal bladder necks. However, after allowing for the influence of body size, and unlike the situation in the bitch, there was no significant difference in proximal urethral length between the two groups. Bladder neck position was significantly related to prostate size (P < 0.001) and it is suggested that this is one reason why castrated male dogs are more prone to urethral sphincter mechanism incompetence than entire animals. A logistic regression analysis revealed that both bladder neck position and castration status were significant risk factors for incontinence and that they appeared to be acting independently of each other.
