Xenopus ribosomal gene enhancers function when inserted inside the gene they enhance.

The ribosomal DNA of Xenopus laevis contains repeated sequence elements in the intergenic spacer region that enhance transcription from the adjacent gene promoter (1,2). Previous work has shown that these RNA polymerase I enhancers influence the target promoter when they are in either orientation, at a distance of several kilobases, and only when they are in cis (3-5). In this work, we further show that enhancer activity is unaffected by inserting the enhancers within the transcription unit whose promoter is being enhanced. In addition, enhancer activity does not interfere with transcription through its sequences. The results suggest that the enhancers act at a point prior to the initiation of transcription and that they are likely to be dispensable once transcription has begun.