Phosphorylation of GENOMES UNCOUPLED 4 Alters Stimulation of Mg Chelatase Activity in Angiosperms.
GENOMES UNCOUPLED 4 (GUN4) is a positive regulator of light-dependent chlorophyll biosynthesis. GUN4 activates Mg chelatase (MgCh) that catalyzes the insertion of an Mg2+ion into protoporphyrin IX. We show that Arabidopsis (Arabidopsis thaliana) GUN4 is phosphorylated at Ser 264 (S264), the penultimate amino acid residue at the C terminus. While GUN4 is preferentially phosphorylated in darkness, phosphorylation is reduced upon accumulation of Mg porphyrins. Expression of a phosphomimicking GUN4(S264D) results in an incomplete complementation of the white gun4-2 null mutant and a chlorotic phenotype comparable to gun4 knockdown mutants. Phosphorylated GUN4 has a reduced stimulatory effect on MgCh in vitro and in vivo but retains its protein stability and tetrapyrrole binding capacity. Analysis of GUN4 found in oxygenic photosynthetic organisms reveals the evolution of a C-terminal extension, which harbors the phosphorylation site of GUN4 expressed in angiosperms. Homologs of GUN4 from Synechocystis and Chlamydomonas lack the conserved phosphorylation site found in a C-terminal extension of angiosperm GUN4. Biochemical studies proved the importance of the C-terminal extension for MgCh stimulation and inactivation of GUN4 by phosphorylation in angiosperms. An additional mechanism regulating MgCh activity is proposed. In conjunction with the dark repression of 5-aminolevulinic acid synthesis, GUN4 phosphorylation minimizes the flow of intermediates into the Mg branch of the tetrapyrrole metabolic pathway for chlorophyll biosynthesis.
Plant Physiology, Institute of Biology, Humboldt-Universität zu Berlin, 10115 Berlin, Germany (A.S.R., C.H., L.H., F.K., B.G.); and.|Computational Biology, Institute for Biology and Biotechnology of Plants, University of Münster, 48143 Münster, Germany (C.F.).
Andreas Sven
0000-0002-2293-7297
AS
Richter
Plant Physiology, Institute of Biology, Humboldt-Universität zu Berlin, 10115 Berlin, Germany (A.S.R., C.H., L.H., F.K., B.G.); and.|Computational Biology, Institute for Biology and Biotechnology of Plants, University of Münster, 48143 Münster, Germany (C.F.).
Caroline
C
Hochheuser
Plant Physiology, Institute of Biology, Humboldt-Universität zu Berlin, 10115 Berlin, Germany (A.S.R., C.H., L.H., F.K., B.G.); and.|Computational Biology, Institute for Biology and Biotechnology of Plants, University of Münster, 48143 Münster, Germany (C.F.).
Christian
C
Fufezan
Plant Physiology, Institute of Biology, Humboldt-Universität zu Berlin, 10115 Berlin, Germany (A.S.R., C.H., L.H., F.K., B.G.); and.|Computational Biology, Institute for Biology and Biotechnology of Plants, University of Münster, 48143 Münster, Germany (C.F.).
Laura
L
Heinze
Plant Physiology, Institute of Biology, Humboldt-Universität zu Berlin, 10115 Berlin, Germany (A.S.R., C.H., L.H., F.K., B.G.); and.|Computational Biology, Institute for Biology and Biotechnology of Plants, University of Münster, 48143 Münster, Germany (C.F.).
Franziska
0000-0002-0374-6671
F
Kuhnert
Plant Physiology, Institute of Biology, Humboldt-Universität zu Berlin, 10115 Berlin, Germany (A.S.R., C.H., L.H., F.K., B.G.); and bernhard.grimm@hu-berlin.de.|Computational Biology, Institute for Biology and Biotechnology of Plants, University of Münster, 48143 Münster, Germany (C.F.) bernhard.grimm@hu-berlin.de.
Bernhard
0000-0002-9730-1074
B
Grimm
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D003624:Darkness; D057075:Enzyme Assays; D020869:Gene Expression Profiling; D055786:Gene Knockout Techniques; D005816:Genetic Complementation Test; D005838:Genotype; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008190:Lyases / Q000378:metabolism*; D008954:Models, Biological; D009154:Mutation / Q000235:genetics; D010084:Oxidation-Reduction; D010641:Phenotype; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D030821:Plants, Genetically Modified; D018087:Plastids / Q000378:metabolism; D011166:Porphyrins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D029681:Arabidopsis Proteins; C472017:GUN4 protein, Arabidopsis; D047908:Intracellular Signaling Peptides and Proteins; D011166:Porphyrins; D011994:Recombinant Proteins; D010768:Phosphoserine; D008190:Lyases; C086788:magnesium chelatase
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