Transcription factor IIIA (
TFIIIA), the canonical zinc-finger protein, is
a protein of relative molecular mass 39,000 (39K) that is required for transcription of 5S-ribosomal subunit genes in Xenopus. It binds in a sequence-specific manner to the internal control region of the 5S gene (see Fig. 1) and facilitates transcription of the gene by
RNA polymerase III. It also binds to the 5S gene product to form a 7S
ribonucleoprotein particle. In oocytes the 7S particle acts as a storage form of the
RNA to be utilized later in development.
TFIIIA binds to
DNA through its 30 K N-terminal domain, which contains nine zinc-fingers.
TFIIIA was the first protein described to have this type of
DNA binding motif, but numerous other
proteins have now been shown to have zinc-finger domains. A structure for a single zinc-finger from the yeast protein ADR1, was recently proposed based on two-dimensional NMR data (ref. 8), and a similar structure was proposed based on comparison with crystal structures of other
metalloproteins. Although models for the interaction of
TFIIIA with the 5S-ribosomal gene
DNA have been proposed, based on nuclease digestion and methylation interference data, little precise structural information is available for
TFIIIA and the physical basis for the interaction of zinc-fingers with
DNA is not understood. Using both circular permutation and circularization assays we provide convincing biochemical evidence that
TFIIIA bends the
DNA at the internal promoter of the 5S gene.