Isolation of transcription factor IIIC from Dictyostelium discoideum

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Abstract
Transcription factor IIIC (TFIIIC) binds in a sequence‐specific manner to RNA‐polymerase‐III‐transcribed genes (e.g. tRNA genes). It sequesters other transcription factors into the preformed complex, thereby activating transcription by RNA polymerase III. The Dictyostelium discoideum homologue of TFIIIC was highly purified by affinity chromatography based on its tDNA‐binding activity. This TFIIIC homologue is a multicomponent factor (molecular mass 380 kDa), which binds to the B‐box element of the internal tRNA gene promoter without significant A‐box interaction. Partially purified D. discoideum TFIIIC is able to functionally complement a human RNA polymerase III in vitro transcription system depleted of human TFIIIC. We provide evidence that partially purified D. discoideum TFIIIC interacts in vitro with gene‐external B‐box elements present downstream of many D. discoideum tRNA genes.