In vivo and in vitro detection of the leader RNA of the histidine operon of Escherichia coli K-12 (attenuation/leader RNA/minicells)

The DNA of the attenuator region of the histidine operon of Escherichia coli has been transcribed in a purified in vitro system and found to synthesize two major RNA transcripts. The first one, 180 nucleotides long, has been identified as the his- tidine-specific leader RNA. It contains the coding sequence for the leader peptide (Di Nocera, P. P., Blasi, F., Di Lauro, R., Frunzio, R. & Bruni, C. B. (1978) Proc. Natl. Acad. Sci. USA 75, 4276-4280) and is terminated at the attenuator site. Termination of transcrip- tion at this site is extremely efficient in the in vitro system. The leader RNA also has been detected in vivo in a minicell producer strain transformed with plasmids harboring the regulatory region of the histidine operon of E. coli. A second RNA molecule is syn- thesized in the in vitro system. It has a divergent direction of tran- scription with respect to the histidine leader RNA, but its role, if any, in the regulation of the histidine operon remains to be as- certained. The existence of the histidine leader RNA lends support to the regulatory mechanism which postulates that regulation of the histidine operon is dependent on the alternative secondary structures that the leader RNA may assume, depending on whether or not the histidine-rich leader peptide is translated.