Characterization of a Euglena gracilis chloroplast RNA polymerase specific for ribosomal RNA genes.

Euglena gracilis chloroplasts contain a 145,000-base pair chromosome that encodes genes for ribosomal, transfer, and messenger RNAs. These genes are transcribed within the organelle by chloroplast RNA polymerase activities that are specific for different classes of RNA. Two transcriptional activities have been isolated from Euglena chloroplasts. (Greenberg, B. M., Narita, J. O., DeLuca-Flaherty, C., Gruissem, W., Rushlow, K. A., and Hallick, R. B. (1984) J. Biol. Chem. 259, 14880-14887). One, the "soluble extract," contains enzymes active in tRNA transcription and processing. The other activity, the transcriptionally active chromosome, consisting of a chloroplast DNA-dependent RNA polymerase tightly bound to chloroplast DNA, only transcribes rRNA genes even though the entire chloroplast genome is present. We have extensively purified the transcriptionally active chromosome using high salt concentrations to dissociate loosely bound proteins. The result is a highly enriched extract containing three major polypeptides of Mr 116,000-118,000, 83,000-88,000, and 24,000-26,000 that retains complete selectivity for rDNA transcription. It is probable that one, or both, of the high molecular weight proteins are functional components of the DNA-dependent RNA polymerase. The identification and characterization of the transcriptionally active chromosome is a first step towards understanding how chloroplast rRNA synthesis is regulated.