Subcellular location of the tetrapyrrole synthesis enzyme porphobilinogen deaminase in higher plants: an immunological investigation

Witty, Michael; Jones, Russell M.; Robb, Matthew S.; Jordan, Peter M.; Smith, Alison G.

doi:10.1007/bf00195187

Cited by 14 publications

(6 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…RUG1 is broadly expressed, as shown by data deposited at different publicly available microarray databases [Genevestigator (https://www.genevestigator.com/gv/) and the BIO-array resource (BAR; http://bar.utoronto.ca/efp/cgi-bin/efpWeb.cgi)] and consistent with other experimental results that detected PBGD in different organs of Arabidopsis [41], [42] and pea [40]. Interestingly, we found that overexpression of Arabidopsis PBGD in a wild-type genetic background leads to the appearance of supernumerary shoot apical meristems and occasionally small necrotic patches in the leaves (Figure S2d–f).…”

Section: Resultssupporting

confidence: 88%

“…In animals and yeast, PBGD is a cytosolic protein but in higher plants and algae, it is targeted to the chloroplast [40]. In Arabidopsis, PBGD is a chloroplast protein encoded by a single-copy gene [41], [42]. The overall sequence similarity between the PBGD of Arabidopsis and other organisms is moderately high: 76, 75, 74, 46, 38, 37, 37 and 35% identity for pea, wheat, rice, Escherichia coli , human, mouse, Danio rerio and Saccharomyces cerevisiae , respectively (Figure 5).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

PORPHOBILINOGEN DEAMINASE Deficiency Alters Vegetative and Reproductive Development and Causes Lesions in Arabidopsis

et al. 2013

View full text Add to dashboard Cite

The Arabidopsis rugosa1 (rug1) mutant has irregularly shaped leaves and reduced growth. In the absence of pathogens, leaves of rug1 plants have spontaneous lesions reminiscent of those seen in lesion-mimic mutants; rug1 plants also express cytological and molecular markers associated with defence against pathogens. These rug1 phenotypes are made stronger by dark/light transitions. The rug1 mutant also has delayed flowering time, upregulation of the floral repressor FLOWERING LOCUS C (FLC) and downregulation of the flowering promoters FT and SOC1/AGL20. Vernalization suppresses the late flowering phenotype of rug1 by repressing FLC. Microarray analysis revealed that 280 nuclear genes are differentially expressed between rug1 and wild type; almost a quarter of these genes are involved in plant defence. In rug1, the auxin response is also affected and several auxin-responsive genes are downregulated. We identified the RUG1 gene by map-based cloning and found that it encodes porphobilinogen deaminase (PBGD), also known as hydroxymethylbilane synthase, an enzyme of the tetrapyrrole biosynthesis pathway, which produces chlorophyll, heme, siroheme and phytochromobilin in plants. PBGD activity is reduced in rug1 plants, which accumulate porphobilinogen. Our results indicate that Arabidopsis PBGD deficiency impairs the porphyrin pathway and triggers constitutive activation of plant defence mechanisms leading to leaf lesions and affecting vegetative and reproductive development.

show abstract

Section: Resultssupporting

confidence: 88%

Section: Resultsmentioning

confidence: 99%

PORPHOBILINOGEN DEAMINASE Deficiency Alters Vegetative and Reproductive Development and Causes Lesions in Arabidopsis

et al. 2013

View full text Add to dashboard Cite

show abstract

“…We therefore used a precursor for another tetrapyrrole biosynthesis enzyme, porphobilinogen deaminase, which had previously been shown to be imported into pea chloroplasts (14) and which had been determined to be located exclusively within the plastids (30). Fig.…”

Section: Origin Of the Additional 38-kda Band After Protease Treatmenmentioning

confidence: 99%

A Single Precursor Protein for Ferrochelatase-I fromArabidopsis Is Imported in Vitro into Both Chloroplasts and Mitochondria

Chow¹,

Singh²,

Roper³

et al. 1997

Journal of Biological Chemistry

View full text Add to dashboard Cite

Ferrochelatase is the last enzyme of heme biosynthesis and in higher plants is found in both chloroplasts and mitochondria. We have isolated cDNAs for two isoforms of ferrochelatase from Arabidopsis thaliana, both of which are imported into isolated chloroplasts. In this paper we show that ferrochelatase-I is also imported into isolated pea mitochondria with approximately the same efficiency as into chloroplasts. Processing of the precursor was observed with both chloroplast stroma and mitochondrial matrix extracts. This was inhibited by EDTA, indicating it was due to the specific processing proteases. The specificity of import was verified by the fact that the mitochondrial preparation did not import the precursor of the light-harvesting chlorophyll a/b protein precursor or the precursor of porphobilinogen deaminase, an earlier enzyme of tetrapyrrole biosynthesis, both of which are exclusively chloroplastlocated. Furthermore, import of ferrochelatase-I precursor into mitochondria was inhibited by valinomycin, but this had no effect on its import into chloroplasts. Thus a single precursor molecule is recognized by the import machinery of the two organelles. The implications for the targeting of ferrochelatase in a possible protective role against photooxidative stress are discussed.

show abstract

“…Protogen oxidase activity has also been measured in plasma membrane [11,12] and endoplasmic reticulum [13], although this activity had different characteristics from those in the plastids and mitochondria. In contrast, all the steps up to and including coproporphyrinogen III oxidase are confined to plastids [9,[14][15][16], and it is likely that protogen IX is exported from these organelles to other subcellular locations. Evidence for the export of this compound was obtained after the incubation of isolated barley plastids with the tetrapyrrole precursor 5-aminolaevulinic acid [17].…”

Section: Introductionmentioning

confidence: 99%

Measurement of ferrochelatase activity using a novel assay suggests that plastids are the major site of haem biosynthesis in both photosynthetic and non-photosynthetic cells of pea (Pisum sativum L.)

Cornah

Roper

Singh

et al. 2002

Biochemical Journal

View full text Add to dashboard Cite

Ferrochelatase is the terminal enzyme of haem biosynthesis, catalysing the insertion of ferrous iron into the macrocycle of protoporphyrin IX, the last common intermediate of haem and chlorophyll synthesis. Its activity has been reported in both plastids and mitochondria of higher plants, but the relative amounts of the enzyme in the two organelles are unknown. Ferrochelatase is difficult to assay since ferrous iron requires strict anaerobic conditions to prevent oxidation, and in photosynthetic tissues chlorophyll interferes with the quantification of the product. Accordingly, we developed a sensitive fluorimetric assay for ferrochelatase that employs Co2+ and deuteroporphyrin in place of the natural substrates, and measures the decrease in deuteroporphyrin fluorescence. A hexane-extraction step to remove chlorophyll is included for green tissue. The assay is linear over a range of chloroplast protein concentrations, with an average specific activity of 0.68nmol·min−1·mg of protein−1, the highest yet reported. The corresponding value for mitochondria is 0.19nmol·min−1·mg of protein−1. The enzyme is inhibited by N-methylprotoporphyrin, with an estimated IC50 value of ≈ 1nM. Using this assay we have quantified ferrochelatase activity in plastids and mitochondria from green pea leaves, etiolated pea leaves and pea roots to determine the relative amounts in the two organelles. We found that, in all three tissues, greater than 90% of the activity was associated with plastids, but ferrochelatase was reproducibly detected in mitochondria, at levels greater than the contaminating plastid marker enzyme, and was latent. Our results indicate that plastids are the major site of haem biosynthesis in higher plant cells, but that mitochondria also have the capacity for haem production.

show abstract

Subcellular location of the tetrapyrrole synthesis enzyme porphobilinogen deaminase in higher plants: an immunological investigation

Cited by 14 publications

References 27 publications

PORPHOBILINOGEN DEAMINASE Deficiency Alters Vegetative and Reproductive Development and Causes Lesions in Arabidopsis

PORPHOBILINOGEN DEAMINASE Deficiency Alters Vegetative and Reproductive Development and Causes Lesions in Arabidopsis

A Single Precursor Protein for Ferrochelatase-I fromArabidopsis Is Imported in Vitro into Both Chloroplasts and Mitochondria

Measurement of ferrochelatase activity using a novel assay suggests that plastids are the major site of haem biosynthesis in both photosynthetic and non-photosynthetic cells of pea (Pisum sativum L.)

Contact Info

Product

Resources

About