Regulation of lysine synthesis in transgenic potato plants expressing a bacterial dihydrodipicolinate synthase in their chloroplasts.
The essential amino acid lysine is synthesized in higher plants by a complex pathway that is predominantly regulated by feedback inhibition of two enzymes, namely aspartate kinase (AK) and dihydrodipicolinate synthase (DHPS). Although DHPS is thought to play a major role in this regulation, the relative importance of AK is not known. In order to study this regulation, we have expressed in the chloroplasts of transgenic potato plants a DHPS derived from Escherichia coli at a level 50-fold above the endogenous DHPS. The bacterial enzyme is much less sensitive to lysine inhibition than its potato counterpart. DHPS activity in leaves, roots and tubers of the transgenic plants was considerably higher and more resistant to lysine inhibition than in control untransformed plants. Furthermore, this activity was accompanied by a significant increase in level of free lysine in all three tissues. Yet, the extent of lysine overproduction in potato leaves was significantly lower than that previously reported in leaves of transgenic plants expressing the same bacterial enzyme, suggesting that in potato, AK may also play a major regulatory role in lysine biosynthesis. Indeed, the elevated level of free lysine in the transgenic potato plants was shown to inhibit the lysine-sensitive AK activity in vivo. Our results support previous reports showing that DHPS is the major rate-limiting enzyme for lysine synthesis in higher plants, but they suggest that additional plant-specific regulatory factors are also involved.
Perl
A
A
Department of Plant Genetics, Weizmann Institute of Science, Rehovot, Israel.
Shaul
O
O
Galili
G
G
D001222:Aspartate Kinase / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002736:Chloroplasts; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D006836:Hydro-Lyases / Q000235:genetics* / Q000378:metabolism; D008239:Lysine / Q000096:biosynthesis* / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012231:Rhizobium / Q000235:genetics; D011198:Solanum tuberosum / Q000235:genetics*; D014170:Transformation, Genetic / Q000235:genetics
D011993:Recombinant Fusion Proteins; D001222:Aspartate Kinase; D006836:Hydro-Lyases; C020040:4-hydroxy-tetrahydrodipicolinate synthase; D008239:Lysine