The promoter of the Arabidopsis thaliana plastocyanin gene contains a far upstream enhancer-like element involved in chloroplast-dependent expression.
Plastocyanin is part of the photosynthetic electron transport chain in the chloroplast and is encoded in the nucleus. Expression of the Arabidopsis thaliana plastocyanin gene is organ specific: high mRNA levels are observed in young green parts of the plant. Furthermore, expression is dependent on the presence of light and functional chloroplasts. When grown in the presence of norflurazon under white light conditions, resulting in the photo-oxidative destruction of the chloroplast, plastocyanin mRNA levels are strongly reduced. A -1579 to -9 promoter fragment confers light-regulated and chloroplast-dependent expression to the beta-glucuronidase reporter gene in transgenic tobacco plants. This suggests that regulation takes place at the level of transcription. A plastocyanin promoter deletion series ranging from -1579 to -121 which was also tested in tobacco, revealed the presence of a strong positive regulating element (PRE) in the -1579 to -705 region. Deletion of this part of the promoter resulted in a approximately 100-fold reduction of GUS expression as measured in mature leaves. Surprisingly, this enhancer-like element was capable of stimulating transcription from a position downstream of its reporter. Moreover, it could also activate a truncated CaMV 35S promoter. Deletion of this element coincides with the loss of chloroplast-dependency of reporter gene expression, as judged by norflurazon treatment of transgenic seedlings. So, the activity of the PRE itself might depend on the presence of functional chloroplasts.
Vorst
O
O
Department of Molecular Cell Biology, University of Utrecht, The Netherlands.
Kock
P
P
Lever
A
A
Weterings
B
B
Weisbeek
P
P
Smeekens
S
S
D017360:Arabidopsis / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D002736:Chloroplasts / Q000378:metabolism*; D003001:Cloning, Molecular; D004247:DNA; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation / Q000528:radiation effects; D017343:Genes, Plant; D008027:Light; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D010970:Plastocyanin / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014026:Nicotiana / Q000235:genetics
D004247:DNA; D010970:Plastocyanin