Molecular Characterisation of the 76 kDa Iron-Sulphur Protein Subunit of Potato Mitochondrial Complex I

Rasmusson, Allan G.; Heiser, Volker; Irrgang, Klaus−Dieter; Brennicke, Axel; Grohmann, Lutz

doi:10.1093/oxfordjournals.pcp.a029380

Cited by 19 publications

(10 citation statements)

References 40 publications

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“…Difference in size between precursor and mature NADH:ubiquinone oxidoreductase could not be resolved in the gel system most likely because of the high molecular mass of the protein (ref. 41; L. Grohmann, personal communication). Protoporphyrinogen IX oxidase I and coproporphyrinogen oxidase were bound to mitochondria but obviously not protected from protease treatment.…”

Section: Resultsmentioning

confidence: 99%

Cloning and characterization of a plastidal and a mitochondrial isoform of tobacco protoporphyrinogen IX oxidase

Lermontová

Kruse²,

Mock³

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

177

167

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Protoporphyrinogen IX oxidase is the last enzyme in the common pathway of heme and chlorophyll synthesis and provides precursor for the mitochondrial and plastidic heme synthesis and the predominant chlorophyll synthesis in plastids. We cloned two different, full-length tobacco cDNA sequences by complementation of the protoporphyrin-IX-accumulating Escherichia coli hemG mutant from heme auxotrophy. The two sequences show similarity to the recently published Arabidopsis PPOX, Bacillus subtilis hemY, and to mammalian sequences encoding protoporphyrinogen IX oxidase. One cDNA sequence encodes a 548-amino acid residues protein with a putative transit sequence of 50 amino acid residues, and the second cDNA encodes a protein of 504 amino acid residues. Both deduced protein sequences share 27.2% identical amino acid residues. The first in vitro translated protoporphyrinogen IX oxidase could be translocated to plastids, and the approximately 53-kDa mature protein was detected in stroma and membrane fraction. The second enzyme was targeted to mitochondria without any detectable reduction in size. Localization of both enzymes in subcellular fractions was immunologically confirmed. Steadystate RNA analysis indicates an almost synchronous expression of both genes during tobacco plant development, greening of young seedlings, and diurnal and circadian growth. The mature plastidal and the mitochondrial isoenzyme were overexpressed in E. coli. Bacterial extracts containing the recombinant mitochondrial enzyme exhibit high protoporphyrinogen IX oxidase activity relative to control strains, whereas the plastidal enzyme could only be expressed as an inactive peptide. The data presented confirm a compartmentalized pathway of tetrapyrrole synthesis with protoporphyrinogen IX oxidase in plastids and mitochondria.

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Section: Resultsmentioning

confidence: 99%

Cloning and characterization of a plastidal and a mitochondrial isoform of tobacco protoporphyrinogen IX oxidase

Lermontová

Kruse²,

Mock³

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

177

167

View full text Add to dashboard Cite

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“…FNR (Guedeney et al,1996;Quiles and Cuello, 1998;Funk et al, 1999;Quiles et al, 2000) and nuclear-encoded homologs of NuoE, F, and G (Rasmusson et al, 1998;Quiles et al, 2003) have been suggested to be possible candidates, but no genetic or biochemical confirmation of their involvement was obtained. An alternative strategy, using a genetic approach based on Arabidopsis mutant screening and chlorophyll fluorescence imaging, has been used in recent years to identify missing Ndh complex subunits (Hashimoto et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

New Subunits NDH-M, -N, and -O, Encoded by Nuclear Genes, Are Essential for Plastid Ndh Complex Functioning in Higher Plants

et al. 2005

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In higher plants, the Ndh complex reduces plastoquinones and is involved in cyclic electron flow around photosystem I, supplying extra-ATP for photosynthesis, particularly under environmental stress conditions. Based on plastid genome sequences, the Ndh complex would contain 11 subunits (NDH-A to -K), but homologies with bacterial complex indicate the probable existence of additional subunits. To identify missing subunits, tobacco (Nicotiana tabacum) NDH-H was His tagged at its N terminus using plastid transformation. A functional Ndh subcomplex was purified by Ni 2þ affinity chromatography and its subunit composition analyzed by mass spectrometry. Five plastid encoded subunits (NDH-A, -H, -I, -J, and -K) were identified as well as three new subunits (NDH-M, -N, and -O) homologous to cyanobacterial and higher plant proteins. Arabidopsis thaliana mutants missing one of these new subunits lack a functional Ndh complex, and NDH-M and NDH-N are not detected in a tobacco transformant lacking the Ndh complex. We discuss the involvement of these three nuclear-encoded subunits in the functional integrity of the plastidial complex.

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“…Antibodies used were against potato NDB1 EF-hand domain (Michalecka et al, 2004), potato NDA1 (Rasmusson and Agius, 2001), Sauromatum guttatum alternative oxidase (Elthon et al, 1989), Arabidopsis (Arabidopsis thaliana) uncoupling protein (Borecky et al, 2001), glyoxysomal malate dehydrogenase (Gietl et al, 1996), and the mitochondrial complex I subunits NAD9 from wheat (Triticum aestivum; Lamattina et al, 1993) and 78-kD protein from Neurospora crassa (Friedrich et al, 1989). The latter detects the 76-kD subunit in potato (Rasmusson et al, 1998). Catalase activity was detected in the presence of Triton X-100 as reported previously (Struglics et al, 1993).…”

Section: Organelle Analysesmentioning

confidence: 99%

A Redox-Mediated Modulation of Stem Bolting in TransgenicNicotiana sylvestrisDifferentially Expressing the External Mitochondrial NADPH Dehydrogenase

et al. 2009

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Cytosolic NADPH can be directly oxidized by a calcium-dependent NADPH dehydrogenase, NDB1, present in the plant mitochondrial electron transport chain. However, little is known regarding the impact of modified cytosolic NADPH reduction levels on growth and metabolism. Nicotiana sylvestris plants overexpressing potato (Solanum tuberosum) NDB1 displayed early bolting, whereas sense suppression of the same gene led to delayed bolting, with consequential changes in flowering time. The phenotype was dependent on light irradiance but not linked to any change in biomass accumulation. Whereas the leaf NADPH/NADP + ratio was unaffected, the stem NADPH/NADP + ratio was altered following the genetic modification and strongly correlated with the bolting phenotype. Metabolic profiling of the stem showed that the NADP(H) change affected relatively few, albeit central, metabolites, including 2-oxoglutarate, glutamate, ascorbate, sugars, and hexose-phosphates. Consistent with the phenotype, the modified NDB1 level also affected the expression of putative floral meristem identity genes of the SQUAMOSA and LEAFY types. Further evidence for involvement of the NADPH redox in stem development was seen in the distinct decrease in the stem apex NADPH/NADP + ratio during bolting. Additionally, the potato NDB1 protein was specifically detected in mitochondria, and a survey of its abundance in major organs revealed that the highest levels are found in green stems. These results thus strongly suggest that NDB1 in the mitochondrial electron transport chain can, by modifying cell redox levels, specifically affect developmental processes.

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Molecular Characterisation of the 76 kDa Iron-Sulphur Protein Subunit of Potato Mitochondrial Complex I

Cited by 19 publications

References 40 publications

Cloning and characterization of a plastidal and a mitochondrial isoform of tobacco protoporphyrinogen IX oxidase

Cloning and characterization of a plastidal and a mitochondrial isoform of tobacco protoporphyrinogen IX oxidase

New Subunits NDH-M, -N, and -O, Encoded by Nuclear Genes, Are Essential for Plastid Ndh Complex Functioning in Higher Plants

A Redox-Mediated Modulation of Stem Bolting in TransgenicNicotiana sylvestrisDifferentially Expressing the External Mitochondrial NADPH Dehydrogenase

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