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Nucleic Acids Research, 1994, Vol. 22, No. 22 4798-4805
© 1994

MOLECULAR BIOLOGY

Sequence organization of the Acanthamoeba rRNA intergenic spacer: identification of transcriptional enhancers

Qin Yang, Michael G. Zwick and Marvin R. Paule*

Department of Biochemistry and Molecular Biology, and The Cell and Molecular Biology Program, Colorado State University Fort Collins, CO 80523, USA

*To whom correpsondence should be addressed

Received June 6, 1994. Revised September 11, 1994. Accepted September 11, 1994.

The primary sequence of the entire 2330 bp intergenic spacer of the A.castellanii ribosomal RNA gene was determined. Repeated sequence elements averaging 140 bp were identified and found to bind a protein required for optimum initiation at the core promoter. These repeated elements were shown to stimulate rRNA transcription by RNA polymerase I in vitro. The repeats inhibited transcription when placed in trans, and stimulated transcription when in cis, in either orientation, but only when upstream of the core promoter. Thus, these repeated elements have characteristics similar to polymerase I enhancers found in higher eukaryotes. The number of rRNA repeats in Acanthamoeba cells was determined to be 24 per haploid genome, the lowest number so far identified in any eukaryote. However, because Acanthamoeba is polyploid, each cell contains approximately 600 rRNA genes.


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