IDENTIFICATION OF A TFIIIA BINDING-SITE ON THE 5' FLANKING REGION OF THE TFIIIA GENE

Xenopus transcription factor MIA (TFIIIA) regulates 5S ribosomal RNA gene transcription in a positive manner by binding to the internal control region (ICR) of the 5S RNA gene. The present study reports the identification of a TFIIIA binding site on the 5′ flanking region of the Xenopus laevls gene that codes for the synthesis of this transcription factor. This TFIIIA binding site (deduced from TFIIIA-dependent alterations In DNase I protection patterns) extends on the 5′ flanking region from about nucleotide -326 to -264 on the non-coding strand and from -331 to -271 on the coding strand. The affinity of TFIIIA for the 5' flanking region of its own gene is less than that for the 5S RNA gene but within the same order of magnitude. A sequence similarity between this newly identified binding site and the 5S gene ICR is the presence of purine-rich tracts. EDTA chelation of TFIIIA inhibits binding to this 5' flanking element indicating, as with the 5S RNA gene, zinc is required for ONA binding specificity. When TFIIIA is bound to 5S RNA in the 7S particle, the protein does not bind to this 5' flanking region, an observation similar to that observed with binding to the 5S RNA gene. These results indicate TFIIIA is using similar nucleic acid binding domain(s) for interaction with the 5S gene, 5S RNA, and the 5' flanking region of its own gene. This upstream DNA region to which TFIIIA binds has been previously shown to cont in a negative and a positive regulatory element for transcription of the TFIIIA gene (Scotto, K.W., Kaulen, H., and Roeder, R.G., 1989, Genes & Develop. 3, 651-662). The present results indicate TFIIIA binds to the negative control element located at -306 to -289 and possibly interacts/interferes with another transcription factor which bings to the positive control element extending from -271 to -253. © 1990 Oxford University Press.