title: Posttranslational influence of NADPH-dependent thioredoxin reductase C on enzymes in tetrapyrrole synthesis.
abstract: The NADPH-dependent thioredoxin reductase C (NTRC) is involved in redox-related regulatory processes in chloroplasts and nonphotosynthetic active plastids. Together with 2-cysteine peroxiredoxin, it forms a two-component peroxide-detoxifying system that acts as a reductant under stress conditions. NTRC stimulates in vitro activity of magnesium protoporphyrin IX monomethylester (MgPMME) cyclase, most likely by scavenging peroxides. Reexamination of tetrapyrrole intermediate levels of the Arabidopsis (Arabidopsis thaliana) knockout ntrc reveals lower magnesium protoporphyrin IX (MgP) and MgPMME steady-state levels, the substrate and the product of MgP methyltransferase (CHLM) preceding MgPMME cyclase, while MgP strongly accumulates in mutant leaves after 5-aminolevulinic acid feeding. The ntrc mutant has a reduced capacity to synthesize 5-aminolevulinic acid and reduced CHLM activity compared with the wild type. Although transcript levels of genes involved in chlorophyll biosynthesis are not significantly altered in 2-week-old ntrc seedlings, the contents of glutamyl-transfer RNA reductase1 (GluTR1) and CHLM are reduced. Bimolecular fluorescence complementation assay confirms a physical interaction of NTRC with GluTR1 and CHLM. While ntrc contains partly oxidized CHLM, the wild type has only reduced CHLM. As NTRC also stimulates CHLM activity in vitro, it is proposed that NTRC has a regulatory impact on the redox status of conserved cysteine residues of CHLM. It is hypothesized that a deficiency of NTRC leads to a lower capacity to reduce cysteine residues of GluTR1 and CHLM, affecting the stability and, thereby, altering the activity in the entire tetrapyrrole synthesis pathway.
authors.affiliation: Humboldt-University Berlin, Institute of Biology, Plant Physiology, 10115 Berlin, Germany.
authors.forename: Andreas S
authors.initials: AS
authors.lastname: Richter
authors.forename: Enrico
authors.initials: E
authors.lastname: Peter
authors.forename: Maxi
authors.initials: M
authors.lastname: Rothbart
authors.forename: Hagen
authors.initials: H
authors.lastname: Schlicke
authors.forename: Jouni
authors.initials: J
authors.lastname: Toivola
authors.forename: Eevi
authors.initials: E
authors.lastname: Rintamäki
authors.forename: Bernhard
authors.initials: B
authors.lastname: Grimm
mesh_terms: D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D002736:Chloroplasts / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D008027:Light; D008780:Methyltransferases; D009249:NADP / Q000235:genetics / Q000378:metabolism; D010084:Oxidation-Reduction; D054464:Peroxiredoxins; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D018087:Plastids / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D011524:Protoporphyrins / Q000235:genetics / Q000378:metabolism; D036226:Seedlings / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D045725:Tetrapyrroles / Q000235:genetics / Q000378:metabolism; D013880:Thioredoxin-Disulfide Reductase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013879:Thioredoxins / Q000235:genetics / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D011524:Protoporphyrins; D045725:Tetrapyrroles; C039629:magnesium protoporphyrin; D013879:Thioredoxins; D009249:NADP; C028025:protoporphyrin IX; D054464:Peroxiredoxins; D013880:Thioredoxin-Disulfide Reductase; D008780:Methyltransferases; C060234:magnesium-protoporphyrin methyltransferase