Published online 18 February 2004
Nucleic Acids Research, 2004, Vol. 32, No. 3 1159-1165
© 2004 Oxford University Press
Overexpression of phage-type RNA polymerase RpoTp in tobacco demonstrates its role in chloroplast transcription by recognizing a distinct promoter type
1 Institute of Biology (Genetics), Humboldt University Berlin, Chausseestrasse 117, D-10115 Berlin, Germany and 2 Waksman Institute, Rutgers, the State University of New Jersey, 190 Frelinghuysen Road, Piscataway, NJ 08854-8020, USA
*To whom correspondence should be addressed at Institute of Biology (Genetics), Humboldt University Berlin, Chausseestrasse 117, D-10115 Berlin, Germany. Tel: +49 30 20938156; Fax: +49 30 20938141; Email: karsten.liere{at}rz.hu-berlin.de
Plant cells possess three DNA-containing compartments, the nucleus, the mitochondria and the plastids. Accordingly, plastid gene regulation is fairly complex. Albeit plastids retained their own genome and prokaryotic-type gene expression system by a plastid-encoded RNA polymerase (PEP), they need a second nuclear-encoded plastid transcription activity, NEP. Candidate genes for putative NEP catalytic subunits have been cloned in Arabidopsis thaliana (AtRpoTp) and Nicotiana sylvestris (NsRpoTp). To provide evidence for RpoTp as a gene encoding a NEP catalytic subunit, we introduced the AtRpoTp and NsRpoTp cDNAs into the tobacco nucleus under the control of the strong constitutive CaMV 35S promoter. Analysis of transcription from NEP and PEP promoters in these transgenic plants using primer extension assays revealed enhanced transcription from typical type I NEP promoters as PatpB-289 in comparison with the wild type. These data provide direct evidence that RpoTp is a catalytic subunit of NEP and involved in recognition of a distinct subset of type I NEP promoters.
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