title: Tobacco Mg protoporphyrin IX methyltransferase is involved in inverse activation of Mg porphyrin and protoheme synthesis.
abstract: Protoporphyrin, a metabolic intermediate of tetrapyrrole biosynthesis, is metabolized by Mg chelatase and ferrochelatase and is directed into the Mg-branch for chlorophyll synthesis and in the Fe-branch for protoheme synthesis respectively. Regulation of the enzyme activities at the beginning of this branchpoint ensures accurate partition of protoporphyrin, but is still not entirely understood. Transgenic tobacco plants were generated that express antisense or sense RNA for inhibited and excessive expression of Mg protoporphyrin methyltransferase (MgPMT) respectively. This enzyme accepts Mg protoporphyrin from Mg chelatase and catalyses the transfer of a methyl group to the carboxyl group of the C13-propionate side chain. Low MgPMT activity is correlated with reduced Mg chelatase activity and a low synthesis rate of 5-aminolevulinate, but with enhanced ferrochelatase activity. In contrast, high MgPMT activity leads to inverse activity profiles: high activities of Mg chelatase and for 5-aminolevulinate synthesis, but reduced activity of ferrochelatase, indicating a direct influence of MgPMT in combination with Mg chelatase on the metabolic flux of ALA and the distribution of protoporphyrin into the branched pathway. The modified enzyme activities in tetrapyrrole biosynthesis in the transgenic plants can be explained with changes of certain corresponding mRNA contents: increased 5-aminolevulinate synthesis and Mg chelatase activity correlate with enhanced transcript levels of the HemA, Gsa, and CHLH gene encoding glutamyl-tRNA reductase, glutamate-1-semialdehyde aminotransferase and a Mg chelatase subunit respectively. It is proposed that reduced and increased MgPMT activity in chloroplasts is communicated to the cytoplasm for modulating transcriptional activities of regulatory enzymes of the pathway.
authors.affiliation: Institut für Biologie/Pflanzenphysiologie, Humboldt-Universität zu Berlin, Philippstr.13, Haus 12, 10115 Berlin, Germany.
authors.forename: Ali E
authors.initials: AE
authors.lastname: Alawady
authors.forename: Bernhard
authors.initials: B
authors.lastname: Grimm
mesh_terms: D002734:Chlorophyll / Q000378:metabolism; D015870:Gene Expression; D006418:Heme / Q000378:metabolism*; D008190:Lyases / Q000235:genetics / Q000378:metabolism; D008780:Methyltransferases / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011524:Protoporphyrins / Q000378:metabolism*; D045725:Tetrapyrroles / Q000096:biosynthesis*; D014026:Nicotiana / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D019076:Transgenes
chemical_list: D011524:Protoporphyrins; D045725:Tetrapyrroles; D002734:Chlorophyll; C039629:magnesium protoporphyrin; D006418:Heme; D008780:Methyltransferases; C060234:magnesium-protoporphyrin methyltransferase; D008190:Lyases; C086788:magnesium chelatase