Nkx2.5 and Nkx2.6, Homologs of Drosophila tinman, Are Required for Development of the Pharynx

Abstract

Transcription of yeast class III genes involves the formation of a transcription initiation complex that comprises RNA polymerase III (Pol III) and the general transcription factors TFIIIB and TFIIIC. Using a genetic screen for positive regulators able to compensate for a deficiency in a promoter element of the SNR6 gene, we isolated the NHP6A and NHP6B genes. Here we show that the high-mobility-group proteins NHP6A and NHP6B are required for the efficient transcription of the SNR6 gene both in vivo and in vitro. The transcripts of wild-type and promoter-defectiveSNR6 genes decreased or became undetectable in annhp6AΔ nhp6BΔ double-mutant strain, and the protection over the TATA box of the wild-type SNR6 gene was lost innhp6AΔ nhp6BΔ cells at 37°C. In vitro, NHP6B specifically stimulated the transcription of SNR6 templates up to fivefold in transcription assays using either cell nuclear extracts from nhp6AΔ nhp6BΔ cells or reconstituted transcription systems. Finally, NHP6B activated SNR6transcription in a TFIIIC-independent assay. These results indicate that besides the general transcription factors TFIIIB and TFIIIC, additional auxilliary factors are required for the optimal transcription of at least some specific Pol III genes.