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. 1989 Mar;1(3):293–300. doi: 10.1105/tpc.1.3.293

Coordinate expression of transcriptionally regulated isocitrate lyase and malate synthase genes in Brassica napus L.

L Comai 1, R A Dietrich 1, D J Maslyar 1, C S Baden 1, J J Harada 1
PMCID: PMC159762  PMID: 2535504

Abstract

We have analyzed the temporal and spatial expression of genes encoding the glycoxylate cycle enzymes isocitrate lyase and malate synthase in Brassica napus L. to determine whether they are coordinately expressed. Both enzymes participate in reactions associated with lipid mobilization in oilseed plant seedlings and are sequestered in a specialized organelle, the glyoxysome. We have identified an isocitrate lyase cDNA clone containing the complete protein coding region. RNA blot and in situ hybridization studies with isocitrate lyase and malate synthase cDNA clones from B. napus showed that the genes exhibit similar expression patterns. The mRNAs begin to accumulate during late embryogeny, reach maximal levels in seedling cotyledons, are not detected at significant amounts in leaves, and are distributed similarly in cotyledons and axes of seedlings. Furthermore, transcription studies with isolated nuclei indicate that the genes are controlled primarily although not exclusively at the transcriptional level. We conclude that glyoxysome biogenesis is regulated in part through the coordinate expression of isocitrate lyase and malate synthase genes.

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Selected References

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  1. Allen R. D., Trelease R. N., Thomas T. L. Regulation of isocitrate lyase gene expression in sunflower. Plant Physiol. 1988 Feb;86(2):527–532. doi: 10.1104/pp.86.2.527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ballance D. J., Turner G. Gene cloning in Aspergillus nidulans: isolation of the isocitrate lyase gene (acuD). Mol Gen Genet. 1986 Feb;202(2):271–275. doi: 10.1007/BF00331649. [DOI] [PubMed] [Google Scholar]
  3. Beach L. R., Spencer D., Randall P. J., Higgins T. J. Transcriptional and post-transcriptional regulation of storage protein gene expression in sulfur-deficient pea seeds. Nucleic Acids Res. 1985 Feb 11;13(3):999–1013. doi: 10.1093/nar/13.3.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berry J. O., Nikolau B. J., Carr J. P., Klessig D. F. Transcriptional and post-transcriptional regulation of ribulose 1,5-bisphosphate carboxylase gene expression in light- and dark-grown amaranth cotyledons. Mol Cell Biol. 1985 Sep;5(9):2238–2246. doi: 10.1128/mcb.5.9.2238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Borst P. How proteins get into microbodies (peroxisomes, glyoxysomes, glycosomes). Biochim Biophys Acta. 1986 May 5;866(4):179–203. doi: 10.1016/0167-4781(86)90044-8. [DOI] [PubMed] [Google Scholar]
  6. Breidenbach R. W., Kahn A., Beevers H. Characterization of glyoxysomes from castor bean endosperm. Plant Physiol. 1968 May;43(5):705–713. doi: 10.1104/pp.43.5.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Britten R. J., Davidson E. H. Gene regulation for higher cells: a theory. Science. 1969 Jul 25;165(3891):349–357. doi: 10.1126/science.165.3891.349. [DOI] [PubMed] [Google Scholar]
  8. Chen E. Y., Seeburg P. H. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. doi: 10.1089/dna.1985.4.165. [DOI] [PubMed] [Google Scholar]
  9. Comai L., Baden C. S., Harada J. J. Deduced sequence of a malate synthase polypeptide encoded by a subclass of the gene family. J Biol Chem. 1989 Feb 15;264(5):2778–2782. [PubMed] [Google Scholar]
  10. Cox K. H., DeLeon D. V., Angerer L. M., Angerer R. C. Detection of mrnas in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Dev Biol. 1984 Feb;101(2):485–502. doi: 10.1016/0012-1606(84)90162-3. [DOI] [PubMed] [Google Scholar]
  11. Darnell J. E., Jr Variety in the level of gene control in eukaryotic cells. Nature. 1982 Jun 3;297(5865):365–371. doi: 10.1038/297365a0. [DOI] [PubMed] [Google Scholar]
  12. Dente L., Cesareni G., Cortese R. pEMBL: a new family of single stranded plasmids. Nucleic Acids Res. 1983 Mar 25;11(6):1645–1655. doi: 10.1093/nar/11.6.1645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dietrich R. A., Maslyar D. J., Heupel R. C., Harada J. J. Spatial patterns of gene expression in Brassica napus seedlings: identification of a cortex-specific gene and localization of mRNAs encoding isocitrate lyase and a polypeptide homologous to proteinases. Plant Cell. 1989 Jan;1(1):73–80. doi: 10.1105/tpc.1.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gould S. J., Keller G. A., Subramani S. Identification of peroxisomal targeting signals located at the carboxy terminus of four peroxisomal proteins. J Cell Biol. 1988 Sep;107(3):897–905. doi: 10.1083/jcb.107.3.897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Groudine M., Peretz M., Weintraub H. Transcriptional regulation of hemoglobin switching in chicken embryos. Mol Cell Biol. 1981 Mar;1(3):281–288. doi: 10.1128/mcb.1.3.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  17. Hutton D., Stumpf P. K. Fat Metabolism in Higher Plants. XXXVII. Characterization of the beta-Oxidation Systems From Maturing and Germinating Castor Bean Seeds. Plant Physiol. 1969 Apr;44(4):508–516. doi: 10.1104/pp.44.4.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lazarow P. B., Fujiki Y. Biogenesis of peroxisomes. Annu Rev Cell Biol. 1985;1:489–530. doi: 10.1146/annurev.cb.01.110185.002421. [DOI] [PubMed] [Google Scholar]
  19. Lincoln J. E., Fischer R. L. Diverse mechanisms for the regulation of ethylene-inducible gene expression. Mol Gen Genet. 1988 Apr;212(1):71–75. doi: 10.1007/BF00322446. [DOI] [PubMed] [Google Scholar]
  20. Luthe D. S., Quatrano R. S. Transcription in Isolated Wheat Nuclei: I. ISOLATION OF NUCLEI AND ELIMINATION OF ENDOGENOUS RIBONUCLEASE ACTIVITY. Plant Physiol. 1980 Feb;65(2):305–308. doi: 10.1104/pp.65.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Luthe D. S., Quatrano R. S. Transcription in Isolated Wheat Nuclei: II. CHARACTERIZATION OF RNA SYNTHESIZED IN VITRO. Plant Physiol. 1980 Feb;65(2):309–313. doi: 10.1104/pp.65.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Matsuoka M., McFadden B. A. Isolation, hyperexpression, and sequencing of the aceA gene encoding isocitrate lyase in Escherichia coli. J Bacteriol. 1988 Oct;170(10):4528–4536. doi: 10.1128/jb.170.10.4528-4536.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Miernyk J. A., Trelease R. N. Control of Enzyme Activities in Cotton Cotyledons during Maturation and Germination : IV. beta-OXIDATION. Plant Physiol. 1981 Feb;67(2):341–346. doi: 10.1104/pp.67.2.341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nguyen T., Zelechowska M., Foster V., Bergmann H., Verma D. P. Primary structure of the soybean nodulin-35 gene encoding uricase II localized in the peroxisomes of uninfected cells of nodules. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5040–5044. doi: 10.1073/pnas.82.15.5040. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Riezman H., Weir E. M., Leaver C. J., Titus D. E., Becker W. M. Regulation of Glyoxysomal Enzymes during Germination of Cucumber: 3. IN VITRO TRANSLATION AND CHARACTERIZATION OF FOUR GLYOXYSOMAL ENZYMES. Plant Physiol. 1980 Jan;65(1):40–46. doi: 10.1104/pp.65.1.40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Roberts L. M., Lord J. M. Synthesis and posttranslational segregation of glyoxysomal isocitrate lyase from castor bean endosperm. Eur J Biochem. 1981 Sep;119(1):43–49. doi: 10.1111/j.1432-1033.1981.tb05574.x. [DOI] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Smith S. M., Leaver C. J. Glyoxysomal Malate Synthase of Cucumber: Molecular Cloning of a cDNA and Regulation of Enzyme Synthesis during Germination. Plant Physiol. 1986 Jul;81(3):762–767. doi: 10.1104/pp.81.3.762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Turley R. B., Trelease R. N. Cottonseed malate synthase : biogenesis in maturing and germinated seeds. Plant Physiol. 1987 Aug;84(4):1350–1356. doi: 10.1104/pp.84.4.1350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Volokita M., Somerville C. R. The primary structure of spinach glycolate oxidase deduced from the DNA sequence of a cDNA clone. J Biol Chem. 1987 Nov 25;262(33):15825–15828. [PubMed] [Google Scholar]
  31. Walling L., Drews G. N., Goldberg R. B. Transcriptional and post-transcriptional regulation of soybean seed protein mRNA levels. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2123–2127. doi: 10.1073/pnas.83.7.2123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weir E. M., Riezman H., Grienenberger J. M., Becker W. M., Leaver C. J. Regulation of glyoxysomal enzymes during germination of cucumber. Temporal changes in translatable mRNAs for isocitrate lyase and malate synthase. Eur J Biochem. 1980 Dec;112(3):469–477. [PubMed] [Google Scholar]

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