Interaction of Xenopus TFIIIC with a 5S RNA gene

Using fractionated Xenopus transcription factors we have identified and characterized a unique protein-DNA complex formed between TFIIIA, TFIIIC and a 5S RNA gene. The formation of this complex was blocked by specific competitor DNAs and by the inactivation of TFIIIC using two different methods. In addition, TFIIIC activity was retained when the complexes were affinity purified using a reversibly immobilized DNA template. The TFIII(A+C)-5S RNA gene complex has a distinct electrophoretic mobility on band-shift gels and a unique DNase I footprint. The characteristic feature of the DNase I footprint is a TFIIIC-dependent extension of the TFIIIA footprint an additional 25 bp toward the 5' end of the gene. This indicates a direct interaction between Xenopus TFIIIC and the template DNA.