Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Dec;84(23):8390–8393. doi: 10.1073/pnas.84.23.8390

Bacterial heme synthesis is required for expression of the leghemoglobin holoprotein but not the apoprotein in soybean root nodules.

M R O'Brian 1, P M Kirshbom 1, R J Maier 1
PMCID: PMC299548  PMID: 3479799

Abstract

In Bradyrhizobium japonicum/soybean symbiosis, the leghemoglobin (legume hemoglobin) apoprotein is a plant product, but the origin of the heme prosthetic group is not known. B. japonicum strain LO505 is a transposon Tn5-induced cytochrome-deficient mutant; it excreted the oxidized heme precursor coproporphyrin III into the growth medium. Mutant strain LO505 was specifically deficient in protoporphyrinogen oxidase (protoporphyrinogen-IX:oxygen oxidoreductase, EC 1.3.3.4) activity, and thus it could not catalyze the penultimate step in heme biosynthesis. Soybean root nodules formed from this mutant did not contain leghemoglobin, but the apoprotein was synthesized nevertheless. Data show that bacterial heme synthesis is required for leghemoglobin expression, but the heme moiety is not essential for apoleghemoglobin synthesis by the plant. Soybean leghemoglobin, therefore, is a product of both the plant and bacterial symbionts.

Full text

PDF
8392

Images in this article

8392Image
on p.8392
8392Image
on p.8392

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Avissar Y. J., Nadler K. D. Stimulation of tetrapyrrole formation in Rhizobium japonicum by restricted aeration. J Bacteriol. 1978 Sep;135(3):782–789. doi: 10.1128/jb.135.3.782-789.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bisseling T., van den Bos R. C., van Kammen A. The effect of ammonium nitrate on the synthesis of nitrogenase and the concentration of leghemoglobin in pea root nodules induced by Rhizobium leguminosarum. Biochim Biophys Acta. 1978 Feb 13;539(1):1–11. doi: 10.1016/0304-4165(78)90115-0. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. CHU T. C., SISTER A A GREEN, CHU E. J. Paper chromatography of methyl esters of porphyrins. J Biol Chem. 1951 Jun;190(2):643–646. [PubMed] [Google Scholar]
  5. Camadro J. M., Urban-Grimal D., Labbe P. A new assay for protoporphyrinogen oxidase - evidence for a total deficiency in that activity in a heme-less mutant of Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1982 Jun 15;106(3):724–730. doi: 10.1016/0006-291x(82)91771-5. [DOI] [PubMed] [Google Scholar]
  6. Doss M., Ulshöfer B. Porphyrin stability as a function of the number of carboxylic acid side chains. Biochim Biophys Acta. 1971 May 18;237(2):356–360. doi: 10.1016/0304-4165(71)90330-8. [DOI] [PubMed] [Google Scholar]
  7. Granick S., Beale S. I. Hemes, chlorophylls, and related compounds: biosynthesis and metabolic regulation. Adv Enzymol Relat Areas Mol Biol. 1978;46:33–203. doi: 10.1002/9780470122914.ch2. [DOI] [PubMed] [Google Scholar]
  8. Guerinot M. L., Chelm B. K. Bacterial delta-aminolevulinic acid synthase activity is not essential for leghemoglobin formation in the soybean/Bradyrhizobium japonicum symbiosis. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1837–1841. doi: 10.1073/pnas.83.6.1837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Leong S. A., Ditta G. S., Helinski D. R. Heme biosynthesis in Rhizobium. Identification of a cloned gene coding for delta-aminolevulinic acid synthetase from Rhizobium meliloti. J Biol Chem. 1982 Aug 10;257(15):8724–8730. [PubMed] [Google Scholar]
  10. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  11. Nadler K. D., Avissar Y. J. Heme Synthesis in Soybean Root Nodules: I. On the Role of Bacteroid delta-Aminolevulinic Acid Synthase and delta-Aminolevulinic Acid Dehydrase in the Synthesis of the Heme of Leghemoglobin. Plant Physiol. 1977 Sep;60(3):433–436. doi: 10.1104/pp.60.3.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Noel K. D., Stacey G., Tandon S. R., Silver L. E., Brill W. J. Rhizobium japonicum mutants defective in symbiotic nitrogen fixation. J Bacteriol. 1982 Oct;152(1):485–494. doi: 10.1128/jb.152.1.485-494.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. O'Brian M. R., Kirshbom P. M., Maier R. J. Tn5-induced cytochrome mutants of Bradyrhizobium japonicum: effects of the mutations on cells grown symbiotically and in culture. J Bacteriol. 1987 Mar;169(3):1089–1094. doi: 10.1128/jb.169.3.1089-1094.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Porra R. J. A rapid spectrophotometric assay for ferrochelatase activity in preparations containing much endogenous hemoglobin and its application to soybean root-nodule preparations. Anal Biochem. 1975 Sep;68(1):289–298. doi: 10.1016/0003-2697(75)90707-1. [DOI] [PubMed] [Google Scholar]
  15. Roessler P. G., Nadler K. D. Effects of iron deficiency on heme biosynthesis in Rhizobium japonicum. J Bacteriol. 1982 Mar;149(3):1021–1026. doi: 10.1128/jb.149.3.1021-1026.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Săsárman A., Desrochers M. Uroporphyrinogen III cosynthase-deficient mutant of Salmonella typhimurium LT2. J Bacteriol. 1976 Dec;128(3):717–721. doi: 10.1128/jb.128.3.717-721.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Săsărman A., Chartrand P., Lavoie M., Tardif D., Proschek R., Lapointe C. Mapping of a new hem gene in Escherichia coli K12. J Gen Microbiol. 1979 Aug;113(2):297–303. doi: 10.1099/00221287-113-2-297. [DOI] [PubMed] [Google Scholar]
  18. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Urban-Grimal D., Labbe-Bois R. Genetic and biochemical characterization of mutants of Saccharomyces cerevisiae blocked in six different steps of heme biosynthesis. Mol Gen Genet. 1981;183(1):85–92. doi: 10.1007/BF00270144. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES