Light-regulated differential expression of pea chloroplast and cytosolic fructose-1,6-bisphosphatases.
The light-regulated differential expression of pea chloroplast and cytosolic fructose-1,6-bisphosphatases (FBPase) was investigated using enzyme activity assay, immunoblot, and Northern blot analyses. The enzyme activities of both chloroplast and cytosolic FBPases gradually increased under continuous white light illumination, although the increase in chloroplast FBPase was more drastic. Northern and immunoblot analyses also indicated that light stimulated the expression of both enzymes. Enzyme activity and the transcript levels of both enzymes gradually decreased under the dark treatment, although protein levels were unchanged for up to 24 h following the initiation of culture in the dark, indicating that reversible modifications of the enzymes may occur during the transition from light to dark (or the reverse). Light pulse experiments using blue (420 nm) and red/far-red (660/730 nm) light were carried out to analyze the photoreceptors related to the light-mediated expression of both enzymes. Expression of the chloroplast enzyme was very sensitive to red or far-red light pulses-it was induced by red light, but suppressed by far-red light pulses, as determined by enzyme activity, immunoblot, and Northern blot analyses, suggesting that red light signaling is involved in the control of chloroplast FBPase expression. However, cytosolic FBPase was virtually insensitive to blue or red/far-red light pulses in terms of enzyme activity, as determined by protein and transcript levels, indicating that cytosolic enzyme expression is not directly regulated by light signals. Instead, the expression of the cytosolic enzyme may be closely related to photosynthetic energy conversion accompanied by continuous white light irradiation.
Lee
S-W
SW
Plant Metabolism Research Center, Kyung Hee University, 449-701 Suwon, Korea.
Hahn
T-R
TR
D015152:Blotting, Northern; D015153:Blotting, Western; D002736:Chloroplasts / Q000201:enzymology* / Q000528:radiation effects; D003600:Cytosol / Q000201:enzymology / Q000528:radiation effects; D003624:Darkness; D006597:Fructose-Bisphosphatase / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000528:radiation effects; D018506:Gene Expression Regulation, Plant / Q000528:radiation effects; D008027:Light; D018532:Pisum sativum / Q000201:enzymology / Q000235:genetics* / Q000528:radiation effects
D006597:Fructose-Bisphosphatase