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. 1981 Jun;78(6):3423–3427. doi: 10.1073/pnas.78.6.3423

Overlapping divergent genes in the maize chloroplast chromosome and in vitro transcription of the gene for tRNAHis

Zsuzsanna Schwarz 1,*, Setsuko O Jolly 1, Andre A Steinmetz 1, Lawrence Bogorad 1
PMCID: PMC319580  PMID: 16593030

Abstract

In the presence of the S polypeptide, maize chloroplast DNA-dependent RNA polymerase preferentially transcribes sequences within the 2200-nucleotide-pair-long maize chloroplast chromosome fragment Eco [unk] from a supercoiled chimeric plasmid cloned in Escherichia coli [Jolly, S. O. & Bogorad, L. (1980) Proc. Natl. Acad. Sci. USA 77, 822-826]. Eco [unk] contains one gene for tRNAHis and one for a 1.6-kilobase RNA that includes an open reading frame. These two genes overlap by at least a few nucleotides and are transcribed divergently from complementary DNA strands. This indicates possible transcriptional regulation of chloroplast DNA at the nucleotide level. The 5′ end of tRNAHis (G-U-G) isolated from maize chloroplasts is indistinguishable from that of the transcript produced from Eco [unk] in vitro by maize chloroplast DNA-dependent RNA polymerase. This purified system initiates RNA synthesis faithfully and exhibits preference for some chloroplast genes. Maize chloroplast DNA for tRNAHis lacks the sequence C-C-A at its 3′ terminus; it is presumably added post-transcriptionally. Maize tRNAHis has both prokaryotic and eukaryotic features.

Keywords: DNA sequence analysis, 1.6-kilobase RNA and tRNAHis genes, S1 nuclease mapping, chloroplast RNA polymerase, transcription initiation site

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