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Volume 272, Number 21, Issue of May 23, 1997 pp. 13676-13682
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Organ-specific Transcription of the rrn Operon in Spinach Plastids

(Received for publication, February 11, 1997, and in revised form, March 19, 1997)

From the Laboratoire de Génétique Moléculaire des Plantes, Université Joseph Fourier and CNRS, B. P. 53, F-38041 Grenoble, France

The spinach rrn operon is used as a model system to study transcriptional regulation in higher plant photosynthetic and non-photosynthetic plastids. We performed capping experiments to determine whether P1, PC, or P2 promoters are employed for rrn transcription start sites in cotyledon and root tissues. By using a new method of analysis of capped RNA we demonstrate for the first time that 1) in both organs the rrn operon is expressed in a constitutive manner by cotranscription with the preceding tRNA(GAC)^Val gene, and 2) the PC transcription start site is used only in cotyledons and leaves, i.e. we demonstrate the organ-specific usage of a plastid promoter. Both start sites, PC and that of the tRNA(GAC)^Val cotranscript, lack Escherichia coli-like consensus sequences. The cotranscript is initiated 457 base pairs upstream of the tRNA(GAC)^Val gene. The PC-specific DNA-binding factor, CDF2, is not detectable in root tissues confirming its regulatory role in PC-initiated rrn expression and the organ specificity of PC expression. Furthermore, our results show that rrn operon expression patterns differ in spinach and tobacco indicating species-specific transcriptional regulation of plant plastid gene expression.

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