ADAMTS16 (adisintegrin and metalloproteinase domain with thrombospondin motifs) is a recently discovered metalloproteinase with so far unknown physiological function. Due to structural similarity with other ADAMTS family members one would predict a role for ADAMTS16 in extracellular matrix degradation and angiogenesis. Recent findings indicate a link between arterial hypertension and ADAMTS16 gene polymorphisms. The aim of this study was to analyze the expression pattern of ADAMTS16 and to identify transcriptional pathways involved in its regulation. Immunohistochemistry and in situ mRNA hybridization (ISH) revealed a co-expression of ADAMTS16 and WT1 in embryonic kidneys and gonads, which require WT1 for normal development. ADAMTS16 mRNA was reduced by 90±3% after WT1 knockdown by siRNA interference in mesonephros-derived M15 cells. Conversely, forced expression of WT1 increased ADAMTS16 transcripts more than 5-fold in human embryonic kidney (HEK) 293 cells. A more than 3-fold stimulation of the ADAMTS16 promoter by WT1 protein was determined in transient co-transfection experiments. Interaction of WT1 protein with the ADAMTS16 promoter in cultured M15-cells was confirmed by chromatin immunoprecipitation (ChIP). In silico analysis and electrophoretic mobility shift assays proved three WT1 binding elements in the ADAMTS16 promoter in close proximity to the transcription start site. Treatment of COV434 human granulosa cells with azadeoxycytidin to inhibit DNA methylation caused a 3.5-fold increase of ADAMTS16 mRNA. In conclusion, expression of the ADAMTS16 gene is stimulated by the WT1 transcription factor and sensitive to DNA methylation. Furthermore, our results suggest an involvement of ADAMTS16 in tissue remodelling in the testis and ovary.