The transcriptional activator Staf, originally identified in Xenopus laevis, is implicated in the enhanced transcription of small nuclear RNA (snRNA) and snRNA-type genes by RNA polymerases II (Pol II) and III (Pol III). This zinc finger protein also possesses the capacity to stimulate expression from a Pol II mRNA promoter. Here, we report a study on two human proteins, ZNF76 and ZNF143, that are 64 and 84% identical to their Xenopus counterpart, respectively. Northern blot analysis revealed that ZNF76 and ZNF143 mRNAs were expressed in all normal adult tissues examined. By using in vivo and in vitro assays, we have analyzed the DNA binding capacities and transcriptional properties of ZNF76 and ZNF143. The binding affinities of ZNF76 and ZNF143 for Staf divergent responsive elements were determined by gel shift assays, which revealed that the two proteins bound a same DNA motif with similar affinities. Also, polypeptide sequences containing the seven zinc fingers of ZNF76 and ZNF143 could efficiently repress in vivo the activated transcription from an snRNA-type promoter. Transfection experiments in Drosophila cells showed that ZNF76 and ZNF143 can activate transcription from an mRNA promoter through the Staf binding site. Finally, chimeric ZNF76 and ZNF143 proteins, carrying a heterologous DNA binding domain, are able to activate a Pol II mRNA promoter and snRNA Pol II and Pol III promoters in Xenopus oocytes, through the heterologous DNA binding site. Taken together, these findings demonstrate that ZNF76 and ZNF143 are two members of a same family of transactivator proteins. ZNF143 constitutes the human ortholog of the Xenopus Staf, and ZNF76 is a novel DNA binding protein related to Staf and ZNF143.