Transcriptional regulation by light and phytohormones of the MGD gene in cucumber.
Monogalactosyldiacylglycerol (MGDG) synthase catalyses formation of MGDG, a major structural lipid of chloroplasts. We have already cloned a cDNA for the synthase from a cucumber cDNA library and shown that expression of this gene is regulated by light and a phytohormone, cytokinin. In the present study, we report the molecular basis for transcriptional regulation by light and cytokinin in detail. First, in terms of the enzyme activity, gradual increases in activity mediated by light and cytokinin treatments were observed. At the same time, however, the changes in the mRNA level showed different profiles, with a transient peak during the early stages of light and cytokinin treatment. The interval between the peak level of mRNA and enzyme activation implies the existence of a post-transcriptional regulatory system. In addition, a genomic clone of MGDG synthase isolated from a cucumber genome library was used for a motif search in databases, and this revealed that putative cis-acting elements for light and phytohormones exist in the 5'-upstream region of the MGD cucumber gene. Detailed analysis of this region for light- and cytokinin-responsive activity was performed using a -90 truncated 35 S minimal promoter/luciferase (LUC) reporter gene. It resulted in high levels of LUC expression in etiolated cucumber cotyledons in response to illumination and cytokinin treatment. These results indicate that this 5'-upstream region is involved in light- and cytokinin-enhanced MGD gene expression, and that light- and cytokinin-responsive enhancements of the MGDG synthase activity are regulated, at least in part, at the level of transcription.
Yamaryo
Y
Y
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta, Yokohama, 226-8501, Japan.
Kanai
D
D
Awai
K
K
Masuda
T
T
Shimada
H
H
Takamiya
K
K
Ohta
H
H
D000225:Adenine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D001593:Benzyl Compounds; D002736:Chloroplasts / Q000201:enzymology; D018553:Cucumis sativus / Q000201:enzymology* / Q000235:genetics*; D016207:Cytokines / Q000494:pharmacology; D005700:Galactosyltransferases / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D007701:Kinetin; D008027:Light; D008156:Luciferases / Q000235:genetics; D011401:Promoter Regions, Genetic; D011687:Purines; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects / Q000528:radiation effects
D001593:Benzyl Compounds; D016207:Cytokines; D011687:Purines; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D008156:Luciferases; D005700:Galactosyltransferases; C109990:1,2-diacylglycerol 3-beta-galactosyltransferase; D000225:Adenine; C480551:benzylaminopurine; D007701:Kinetin