GENE DOSAGE AS A POSSIBLE MAJOR DETERMINANT FOR EQUAL EXPRESSION LEVELS OF GENES ENCODING RNA-POLYMERASE SUBUNITS IN THE HYPOTRICHOUS CILIATE EUPLOTES-OCTOCARINATUS

Ciliated protozoa harbor two different types of nuclei in each cell. The diplold micronucleus is the transcriptionally inactive generative nucleus, while the macronuclous contains a highly amplified transcriptionally active genome of lower complexity. The macronuclear genes encoding the two largest subunits of both RNA polymerases I and II of Euplotes octocarinatus were identified by a novel method of two step PCR walking, employing primer pairs derived from telomeric sequences of the organism and known conserved RNA polymerase polypeptide sequences, respectively. The relative gene dosage was determined. The genes are present in equal copy numbers for the respective matching subunits. Northern hybridizations showed comparable amounts of transcripts, as well, within the matching pairs. Mapping of the 5'-termini of the transcripts of the gene sized chromosomes showed that the upstream nontranscribed regions are very short and contain characterisitic sequence motifs which could be the determinants of equal promoter strengths for subunits of a common RNA polymerase.