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. 1997 Jun;114(2):455–466. doi: 10.1104/pp.114.2.455

Differential expression of the multigene family encoding the soybean mitochondrial alternative oxidase.

P M Finnegan 1, J Whelan 1, A H Millar 1, Q Zhang 1, M K Smith 1, J T Wiskich 1, D A Day 1
PMCID: PMC158325  PMID: 9193084

Abstract

The alternative oxidase (AOX) of the soybean (Glycine max L.) inner mitochondrial membrane is encoded by a multigene family (Aox) with three known members. Here, the Aox2 and Aox3 primary translation products, deduced for cDNA analysis, were found to be 38.1 and 36.4 Kd, respectively. Direct N-terminal sequencing of partially purified AOX from cotyledons demonstrates that the mature proteins are 31.8 and 31.6 KD, respectively, implying that processing occurs upon import of these proteins into the mitochondrion. Sequence comparisons show that the processing of plant AOX proteins occurs at a characteristic site and that the AOX2 and AOX3 proteins are more similar to one another than to other AOX proteins, including soybean AOX1. Transcript analysis using a polymerase chain reaction-based assay in conjunction with immunoblot experiments indicates that soybean Aox genes are differentially expressed in a tissue-dependent manner. Moreover, the relative abundance of both Aox2 transcripts and protein in cotyledons increase upon greening of dark-grown seedlings. These results comprehensively explain the multiple AOX-banding patterns observed on immunoblots of mitochondrial proteins isolated from various soybean tissues by matching protein bands with gene products.

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Selected References

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  1. Berthold D. A., Siedow J. N. Partial purification of the cyanide-resistant alternative oxidase of skunk cabbage (Symplocarpus foetidus) mitochondria. Plant Physiol. 1993 Jan;101(1):113–119. doi: 10.1104/pp.101.1.113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chaudhuri M., Hill G. C. Cloning, sequencing, and functional activity of the Trypanosoma brucei brucei alternative oxidase. Mol Biochem Parasitol. 1996 Dec 2;83(1):125–129. doi: 10.1016/s0166-6851(96)02754-5. [DOI] [PubMed] [Google Scholar]
  3. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  4. Conley C. A., Hanson M. R. Tissue-Specific Protein Expression in Plant Mitochondria. Plant Cell. 1994 Jan;6(1):85–91. doi: 10.1105/tpc.6.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cruz-Hernández A., Gómez-Lim M. A. Alternative oxidase from mango (Mangifera indica, L.) is differentially regulated during fruit ripening. Planta. 1995;197(4):569–576. doi: 10.1007/BF00191562. [DOI] [PubMed] [Google Scholar]
  6. Day D. A., Millar A. H., Wiskich J. T., Whelan J. Regulation of Alternative Oxidase Activity by Pyruvate in Soybean Mitochondria. Plant Physiol. 1994 Dec;106(4):1421–1427. doi: 10.1104/pp.106.4.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Day D. A., Wiskich J. T. Regulation of alternative oxidase activity in higher plants. J Bioenerg Biomembr. 1995 Aug;27(4):379–385. doi: 10.1007/BF02110000. [DOI] [PubMed] [Google Scholar]
  8. Elthon T. E., McIntosh L. Characterization and Solubilization of the Alternative Oxidase of Sauromatum guttatum Mitochondria. Plant Physiol. 1986 Sep;82(1):1–6. doi: 10.1104/pp.82.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Elthon T. E., McIntosh L. Identification of the alternative terminal oxidase of higher plant mitochondria. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8399–8403. doi: 10.1073/pnas.84.23.8399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Elthon T. E., Nickels R. L., McIntosh L. Monoclonal antibodies to the alternative oxidase of higher plant mitochondria. Plant Physiol. 1989 Apr;89(4):1311–1317. doi: 10.1104/pp.89.4.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hiser C., Kapranov P., McIntosh L. Genetic modification of respiratory capacity in potato. Plant Physiol. 1996 Jan;110(1):277–286. doi: 10.1104/pp.110.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hiser C., McIntosh L. Alternative Oxidase of Potato Is an Integral Membrane Protein Synthesized de Novo during Aging of Tuber Slices. Plant Physiol. 1990 May;93(1):312–318. doi: 10.1104/pp.93.1.312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kapulnik Y., Yalpani N., Raskin I. Salicylic Acid induces cyanide-resistant respiration in tobacco cell-suspension cultures. Plant Physiol. 1992 Dec;100(4):1921–1926. doi: 10.1104/pp.100.4.1921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kearns A., Whelan J., Young S., Elthon T. E., Day D. A. Tissue-specific expression of the alternative oxidase in soybean and siratro. Plant Physiol. 1992 Jun;99(2):712–717. doi: 10.1104/pp.99.2.712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kumar A. M., Söll D. Arabidopsis alternative oxidase sustains Escherichia coli respiration. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10842–10846. doi: 10.1073/pnas.89.22.10842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Li Q., Ritzel R. G., McLean L. L., McIntosh L., Ko T., Bertrand H., Nargang F. E. Cloning and analysis of the alternative oxidase gene of Neurospora crassa. Genetics. 1996 Jan;142(1):129–140. doi: 10.1093/genetics/142.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McIntosh L. Molecular biology of the alternative oxidase. Plant Physiol. 1994 Jul;105(3):781–786. doi: 10.1104/pp.105.3.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Moore A. L., Siedow J. N. The regulation and nature of the cyanide-resistant alternative oxidase of plant mitochondria. Biochim Biophys Acta. 1991 Aug 23;1059(2):121–140. doi: 10.1016/s0005-2728(05)80197-5. [DOI] [PubMed] [Google Scholar]
  20. Rhoads D. M., McIntosh L. Isolation and characterization of a cDNA clone encoding an alternative oxidase protein of Sauromatum guttatum (Schott). Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2122–2126. doi: 10.1073/pnas.88.6.2122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rhoads D. M., McIntosh L. Salicylic Acid Regulation of Respiration in Higher Plants: Alternative Oxidase Expression. Plant Cell. 1992 Sep;4(9):1131–1139. doi: 10.1105/tpc.4.9.1131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rhoads D. M., McIntosh L. The salicylic acid-inducible alternative oxidase gene aox1 and genes encoding pathogenesis-related proteins share regions of sequence similarity in their promoters. Plant Mol Biol. 1993 Feb;21(4):615–624. doi: 10.1007/BF00014545. [DOI] [PubMed] [Google Scholar]
  23. Ribas-Carbo M., Wiskich J. T., Berry J. A., Siedow J. N. Ubiquinone redox behavior in plant mitochondria during electron transport. Arch Biochem Biophys. 1995 Feb 20;317(1):156–160. doi: 10.1006/abbi.1995.1148. [DOI] [PubMed] [Google Scholar]
  24. Robinson S. A., Yakir D., Ribas-Carbo M., Giles L., Osmond C. B., Siedow J. N., Berry J. A. Measurements of the Engagement of Cyanide-Resistant Respiration in the Crassulacean Acid Metabolism Plant Kalanchoë daigremontiana with the Use of On-Line Oxygen Isotope Discrimination. Plant Physiol. 1992 Nov;100(3):1087–1091. doi: 10.1104/pp.100.3.1087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sakajo S., Minagawa N., Komiyama T., Yoshimoto A. Molecular cloning of cDNA for antimycin A-inducible mRNA and its role in cyanide-resistant respiration in Hansenula anomala. Biochim Biophys Acta. 1991 Aug 27;1090(1):102–108. doi: 10.1016/0167-4781(91)90043-l. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Umbach A. L., Siedow J. N. Covalent and Noncovalent Dimers of the Cyanide-Resistant Alternative Oxidase Protein in Higher Plant Mitochondria and Their Relationship to Enzyme Activity. Plant Physiol. 1993 Nov;103(3):845–854. doi: 10.1104/pp.103.3.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Umbach A. L., Siedow J. N. The reaction of the soybean cotyledon mitochondrial cyanide-resistant oxidase with sulfhydryl reagents suggests that alpha-keto acid activation involves the formation of a thiohemiacetal. J Biol Chem. 1996 Oct 4;271(40):25019–25026. doi: 10.1074/jbc.271.40.25019. [DOI] [PubMed] [Google Scholar]
  29. Vanlerberghe G. C., McIntosh L. Coordinate regulation of cytochrome and alternative pathway respiration in tobacco. Plant Physiol. 1992 Dec;100(4):1846–1851. doi: 10.1104/pp.100.4.1846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Vanlerberghe G. C., McIntosh L. Mitochondrial electron transport regulation of nuclear gene expression. Studies with the alternative oxidase gene of tobacco. Plant Physiol. 1994 Jul;105(3):867–874. doi: 10.1104/pp.105.3.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vanlerberghe G. C., McLntosh L. Signals Regulating the Expression of the Nuclear Gene Encoding Alternative Oxidase of Plant Mitochondria. Plant Physiol. 1996 Jun;111(2):589–595. doi: 10.1104/pp.111.2.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wagner A. M. A role for active oxygen species as second messengers in the induction of alternative oxidase gene expression in Petunia hybrida cells. FEBS Lett. 1995 Jul 17;368(2):339–342. doi: 10.1016/0014-5793(95)00688-6. [DOI] [PubMed] [Google Scholar]
  33. Whelan J., Hugosson M., Glaser E., Day D. A. Studies on the import and processing of the alternative oxidase precursor by isolated soybean mitochondria. Plant Mol Biol. 1995 Feb;27(4):769–778. doi: 10.1007/BF00020229. [DOI] [PubMed] [Google Scholar]
  34. Whelan J., McIntosh L., Day D. A. Sequencing of a soybean alternative oxidase cDNA clone. Plant Physiol. 1993 Dec;103(4):1481–1481. doi: 10.1104/pp.103.4.1481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Whelan J., Millar A. H., Day D. A. The alternative oxidase is encoded in a multigene family in soybean. Planta. 1996;198(2):197–201. doi: 10.1007/BF00206244. [DOI] [PubMed] [Google Scholar]

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