Plant Molecular Biology

, Volume 60, Issue 1, pp 95–106 | Cite as

The Maize Oil Yellow1 (Oy1) Gene Encodes the I Subunit of Magnesium Chelatase

  • Ruairidh J. H. Sawers
  • Joanne Viney
  • Phyllis R. Farmer
  • Rhiannon R. Bussey
  • Gregory Olsefski
  • Katya Anufrikova
  • C. Neil Hunter
  • Thomas P. BrutnellEmail author


Semi-dominant Oil yellow1 (Oy1) mutants of maize (Zea mays) are deficient in the conversion of protoporphyrin IX to magnesium protoporphyrin IX, the first committed step of chlorophyll biosynthesis. Using a candidate gene approach, a cDNA clone was isolated that was predicted to encode the I subunit of magnesium chelatase (ZmCHLI) and mapped to the same genetic interval as Oy1. Allelic variation was identified at ZmCHLI between wild-type plants and plants carrying semi-dominant alleles of Oy1. These differences revealed putative amino acid substitutions that could account for the alterations in protein function. Candidate lesions were tested by introduction of homologous changes into the Synechocystis magnesium chelatase I gene (SschlI) and characterization of the activity of mutant protein variants in an in vitro enzyme activity assay. The results of these analyses suggest that SsChlI protein variants containing the substitutions identified in the dominant Oy1 maize alleles lack activity necessary for magnesium chelation and confer a semi-dominant phenotype via competitive inhibition of wild-type SsChlI.


AAA chelatase magnesium maize oil yellow1 semi-dominance 



ATPases associated with various cellular activities

Fe chelatase


Mg chelatase

magnesium chelatase

Mg proto IX

Mg protoporphyrin IX

proto IX

protoporphyrin IX


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  1. Bochtler, M., Hartmann, C., Song, H.K., Bourenkov, G.P., Bartunik, H.D., Huber, R. 2000The structures of HsIU and the ATP-dependent protease HsIU–HsIVNature403800805CrossRefPubMedGoogle Scholar
  2. Cornah, J.E., Terry, M.J., Smith, A.G. 2003Green or red: what stops the traffic in the tetrapyrrole pathway?Trends Plant Sci.8224230CrossRefPubMedGoogle Scholar
  3. Davey, M.J., Jeruzalmi, D., Kuriyan, J., O’Donnell, M. 2002Motors and switches: AAA+ machines within the replisomeNat. Rev. Mol. Cell. Biol.3826835CrossRefPubMedGoogle Scholar
  4. Davison, P.A., Schubert, H.L., Reid, J.D., Lorg, C., Heroux, A., Hill, C.P., Hunter, C.N. 2005Structural and biochemical characterization of GUN4 suggests a mechanism for its role in chlorophyll biosynthesisBiochemistry4476037612CrossRefPubMedGoogle Scholar
  5. Dooner, H.K. 2002Extensive interallelic polymorphisms drive meiotic recombination into a crossover pathwayPlant Cell1411731183CrossRefPubMedGoogle Scholar
  6. Emanuelson, O., Nielsen, H., Brunak, S., Heijne, G. 2000Predicting subcellular localization of protiens based on their N-terminal amino acid sequenceJ. Mol. Biol.30010051016Google Scholar
  7. Fodje, M.N., Hansson, A., Hansson, M., Olsen, J.G., Gough, S., Willows, R.D., Al-Karadaghi, S. 2001Interplay between an AAA module and an integrin I domain may regulate the function of magnesium chelataseJ. Mol. Biol.311111122CrossRefPubMedGoogle Scholar
  8. Gibson, L.C., Jensen, P.E., Hunter, C.N. 1999Magnesium chelatase from Rhodobacter sphaeroides: initial characterization of the enzyme using purified subunits and evidence for a BchI-BchD complexBiochem. J.337243251CrossRefPubMedGoogle Scholar
  9. Gibson, L.C., Marrison, J.L., Leech, R.M., Jensen, P.E., Bassham, D.C., Gibson, M., Hunter, C.N. 1996A putative Mg chelatase subunit from Arabidopsis thaliana cv C24. Sequence and transcript analysis of the gene, import of the protein into chloroplasts, and in situ localization of the transcript and proteinPlant Physiol.1116171CrossRefPubMedGoogle Scholar
  10. Gibson, L.C., Willows, R.D., Kannangara, C.G., Wettstein, D., Hunter, C.N. 1995Magnesium-protoporphyrin chelatase of Rhodobacter sphaeroides: reconstitution of activity by combining the products of the bchH, -I, and -D genes expressed in Escherichia coliProc. Natl. Acad. Sci. USA9219411944PubMedGoogle Scholar
  11. Grimm B. 2003. Regulatory mechanisms of eukaryotic tetrapyrrole biosynthesis. In: K.M. Kadish, Smith K.M. Smith and R. Guilard (Eds.), The Porphyrin Handbook, New York pp. 1–32.Google Scholar
  12. Hansson, A., Kannangara, C.G., Wettstein, D., Hansson, M. 1999Molecular basis for semidominance of missense mutations in the XANTHA-H (42-kDa) subunit of magnesium chelataseProc. Natl. Acad. Sci. USA9617441749PubMedGoogle Scholar
  13. Hansson, A., Willows, R.D., Roberts, T.H., Hansson, M. 2002Three semidominant barley mutants with single amino acid substitutions in the smallest magnesium chelatase subunit form defective AAA+ hexamersProc. Natl. Acad. Sci. USA991394413949PubMedGoogle Scholar
  14. Hudson, A., Carpenter, R., Doyle, S., Coen, E.S. 1993OLIVE: a key gene required for chlorophyll biosynthesis in Antirrhinum majusEmbo J.1237113719PubMedGoogle Scholar
  15. Iyer, L.M., Leipe, D.D., Koonin, E.V., Aravind, L. 2004Evolutionary history and higher order classification of AAA+ ATPasesJ. Struct. Biol.1461131CrossRefPubMedGoogle Scholar
  16. Jensen, P.E., Gibson, L.C., Henningsen, K.W., Hunter, C.N. 1996Expression of the chlI, chlD, and chlH genes from the Cyanobacterium Synechocystis PCC6803 in Escherichia coli and demonstration that the three cognate proteins are required for magnesium-protoporphyrin chelatase activityJ. Biol. Chem.2711666216667PubMedGoogle Scholar
  17. Jensen, P.E., Gibson, L.C., Hunter, C.N. 1998Determinants of catalytic activity with the use of purified I, D and H subunits of the magnesium protoporphyrin IX chelatase from Synechocystis PCC6803Biochem. J.334335344PubMedGoogle Scholar
  18. Jensen, P.E., Gibson, L.C., Hunter, C.N. 1999ATPase activity associated with the magnesium-protoporphyrin IX chelatase enzyme of Synechocystis PCC6803: evidence for ATP hydrolysis during Mg2+ insertion, and the MgATP-dependent interaction of the ChlI and ChlD subunitsBiochem. J.339127134CrossRefPubMedGoogle Scholar
  19. Jensen, P.E., Willows, R.D., Petersen, B.L., Vothknecht, U.C., Stummann, B.M., Kannangara, C.G., Wettstein, D., Henningsen, K.W. 1996Structural genes for Mg-chelatase subunits in barley: Xantha-f, -g and -hMol. Gen. Genet.250383394PubMedGoogle Scholar
  20. Kannangara, C.G., Vothknecht, U.C., Hansson, M., Wettstein, D. 1997Magnesium chelatase: association with ribosomes and mutant complementation studies identify barley subunit XANTHA-G as a functional counterpart of Rhodobacter subunit BchDMol. Gen. Genet.2548592PubMedGoogle Scholar
  21. Karger, G.A., Reid, J.D., Hunter, C.N. 2001Characterization of the binding of deuteroporphyrin IX to the magnesium chelatase H subunit and spectroscopic properties of the complexBiochemistry4092919299CrossRefPubMedGoogle Scholar
  22. Kim, K.I., Cheong, G.W., Park, S.C., Ha, J.S., Woo, K.M., Choi, S.J., Chung, C.H. 2000Heptameric ring structure of the heat-shock protein ClpB, a protein-activated ATPase in Escherichia coliJ. Mol. Biol.303655666PubMedGoogle Scholar
  23. Kjemtrup, S., Sampson, K.S., Peele, C.G., Nguyen, L.V., Conkling, M.A., Thompson, W.F., Robertson, D. 1998Gene silencing from plant DNA carried by a geminivirusPlant J.1491100CrossRefPubMedGoogle Scholar
  24. Koncz, C., Mayerhofer, R., Koncz-Kalman, Z., Nawrath, C., Reiss, B., Redei, G.P., Schell, J. 1990Isolation of a gene encoding a novel chloroplast protein by T-DNA tagging in Arabidopsis thalianaEmbo J.913371346PubMedGoogle Scholar
  25. Larkin, R.M., Alonso, J.M., Ecker, J.R., Chory, J. 2003GUN4, a regulator of chlorophyll synthesis and intracellular signalingScience299902906CrossRefPubMedGoogle Scholar
  26. Lecerof, D., Fodje, M.N., Alvarez Leon, R., Olsson, U., Hansson, A., Sigfridsson, E., Ryde, U., Hansson, M., Al-Karadaghi, S 2003Metal binding to Bacillus subtilis ferrochelatase and interaction between metal sitesJ. Biol. Inorg. Chem.8452458PubMedGoogle Scholar
  27. Lindstrom, E.W. 1935Some new mutants in maizeIowa State Coll. J. Sci.9451459Google Scholar
  28. Mascia, P.N. 1978Genetic studies of luteus mutantsMNL525658Google Scholar
  29. Mascia, P.N., Robertson, D.S. 1980Genetic studies of the chlorophyll biosynthetic mutants of maizeJ. Hered.711924Google Scholar
  30. Matz, E.C., Burr, F., Burr, B. 1992RFLP mapping of mutantsMNL66107Google Scholar
  31. Mochizuki, N., Brusslan, J.A., Larkin, R., Nagatani, A., Chory, J. 2001Arabidopsis genomes uncoupled 5 (gun5) mutant reveals the involvement of Mg-chelatase H subunit in plastid-to-nucleus signal transductionProc. Natl. Acad. Sci. USA9820532058PubMedGoogle Scholar
  32. Neuffer, M.G., England, D.J. 1995Induced mutations with confirmed locationsMNL694346Google Scholar
  33. Nguyen, L.V. 1995. Transposon Tagging and Isolation of the SULFUR Gene in Tobacco (Nicotiana tabacum). North Carolina State University.Google Scholar
  34. Ogura, T., Wilkinson, A.J. 2001AAA+ superfamily ATPases: common structure-diverse functionGenes Cells6575597CrossRefPubMedGoogle Scholar
  35. Palmer, L.E., Rabinowicz, P.D., O’Shaughnessy, A.L., Balija, V.S., Nascimento, L.U., Dike, S., la Bastide, M., Martienssen, R.A., McCombie, W.R. 2003Maize genome sequencing by methylation filtrationScience30221152117CrossRefPubMedGoogle Scholar
  36. Reid, J.D., Hunter, C.N. 2004Magnesium-dependent ATPase activity and cooperativity of magnesium chelatase from Synechocystis sp. PCC6803J. Biol. Chem.2792689326899PubMedGoogle Scholar
  37. Reid, J.D., Siebert, C.A., Bullough, P.A., Hunter, C.N. 2003The ATPase activity of the ChlI subunit of magnesium chelatase and formation of a heptameric AAA+ ringBiochemistry4269126920PubMedGoogle Scholar
  38. Sawers, R.J., Linley, P.J., Gutierrez-Marcos, J.F., Delli-Bovi, T., Farmer, P.R., Kohchi, T., Terry, M.J., Brutnell, T.P. 2004The Elm1 (ZmHy2) gene of maize encodes a phytochromobilin synthasePlant Physiol.13627712781CrossRefPubMedGoogle Scholar
  39. St Johnston, D. 2002The art and design of genetic screens: Drosophila melanogasterNat. Rev. Genet.3176188CrossRefPubMedGoogle Scholar
  40. Strand, A. 2004Plastid-to-nucleus signallingCurr. Opin. Plant Biol.7621625CrossRefPubMedGoogle Scholar
  41. Strand, A., Asami, T., Alonso, J., Ecker, J.R., Chory, J. 2003Chloroplast to nucleus communication triggered by accumulation of Mg-protoporphyrinIXNature4217983CrossRefPubMedGoogle Scholar
  42. Tenaillon, M.I., Sawkins, M.C., Anderson, L.K., Stack, S.M., Doebley, J., Gaut, B.S. 2002Patterns of diversity and recombination along chromosome 1 of maize (Zea mays ssp. mays L.)Genetics16214011413PubMedGoogle Scholar
  43. Thornsberry, J.M., Goodman, M.M., Doebley, J., Kresovich, S., Nielsen, D., Buckler, E.S. 2001Dwarf8 polymorphisms associate with variation in flowering timeNat. Genet.28286289CrossRefPubMedGoogle Scholar
  44. Vale, R.D. 2000AAA proteins. Lords of the ringJ. Cell. Biol.150F13F19CrossRefPubMedGoogle Scholar
  45. Walker, C.J., Willows, R.D. 1997Mechanism and regulation of Mg-chelataseBiochem J.327321333PubMedGoogle Scholar
  46. Whitelaw, C.A., Barbazuk, W.B., Pertea, G., Chan, A.P., Cheung, F., Lee, Y., Zheng, L., Heeringen, S., Karamycheva, S., Bennetzen, J.L., SanMiguel, P., Lakey, N., Bedell, J., Yuan, Y., Budiman, M.A., Resnick, A., Aken, S., Utterback, T., Riedmuller, S., Williams, M., Feldblyum, T., Schubert, K., Beachy, R., Fraser, C.M., Quackenbush, J. 2003Enrichment of gene-coding sequences in maize by genome filtrationScience30221182120CrossRefPubMedGoogle Scholar
  47. Whitt, S.R., Wilson, L.M., Tenaillon, M.I., Gaut, B.S., Buckler, E.S. 2002Genetic diversity and selection in the maize starch pathwayProc. Natl. Acad. Sci. USA991295912962CrossRefPubMedGoogle Scholar
  48. Wilde, A., Mikolajczyka, S., Alawady, A., Lokstein, H., Grimm, B. 2004The gun4 gene is essential for cyanobacterial porphyrin metabolismFEBS Lett.5119123Google Scholar
  49. Willows, R.D. 2003Biosynthesis of chlorophylls from protoporphyrin IXNat. Prod. Rep.20327341CrossRefPubMedGoogle Scholar
  50. Wilson,  2004Dissection of maize kernel composition and starch production by candidate gene associationPlant Cell1627192733CrossRefPubMedGoogle Scholar

Copyright information

© Springer 2006

Authors and Affiliations

  • Ruairidh J. H. Sawers
    • 1
  • Joanne Viney
    • 2
  • Phyllis R. Farmer
    • 1
  • Rhiannon R. Bussey
    • 3
  • Gregory Olsefski
    • 3
  • Katya Anufrikova
    • 1
  • C. Neil Hunter
    • 2
  • Thomas P. Brutnell
    • 1
    Email author
  1. 1.Boyce Thompson InstituteCornell UniversityIthacaUSA
  2. 2.Department of Molecular Biology and BiotechnologyUniversity of SheffieldSheffieldUK
  3. 3.Cornell UniversityIthacaUSA

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