Spinach cytosolic fructose-1,6-bisphosphatase: I. Its organ-specific and developmental expression characteristics.
Spinach (Spinacia oleracea) cytosolic fructose-1,6-bisphosphatase (FBPase) was purified and the final preparation of protein has a specific activity of about 45 units/mg protein and a single band of molecular mass of 39 kDa. Polyclonal antibody against the protein did not cross-react with chloroplast FBPase, but showed strong cross-reactivity with all plant cytosolic FBPases tested. Studies of the FBPase expression characteristics at early stages of development demonstrated that it was controlled at both the transcriptional and translational levels, and its mRNA was detected even in etiolated cotyledons. This suggests that the expression is not light-inducible. A single transcript was detected in all spinach tissues tested. Western blot analysis revealed two protein bands in the etiolated cotyledons: one was the same size as that present in the mature leaf, and the other slightly smaller. A high enzyme activity was detected in etiolated cotyledons, especially compared to protein levels in Western blots. Expression of the cytosolic FBPase gene during leaf development showed no change in the steady-state level of mRNA, but the protein level and enzyme activity were higher in mature leaves than in young ones, suggesting that the increase in FBPase activity during development is due to an increase in protein synthesis. Young roots showed low enzyme activity, but an unexpectedly high activity was detected in old fiber roots.
Hur
Y
Y
Department of Biology, Chungnam National University, Taejeon, Korea. ykhur@hanbat.chungnam.ac.kr
Vasconcelos
A C
AC
D000906:Antibodies / Q000378:metabolism; D018548:Cotyledon / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D006597:Fructose-Bisphosphatase / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D015870:Gene Expression / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics; D018546:Hypocotyl / Q000201:enzymology; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D018517:Plant Roots / Q000201:enzymology; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D018724:Spinacia oleracea / Q000201:enzymology* / Q000254:growth & development / Q000648:ultrastructure*; D013997:Time Factors; D014018:Tissue Distribution
D000906:Antibodies; D012333:RNA, Messenger; D006597:Fructose-Bisphosphatase