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. 1989 Jun;86(12):4435–4439. doi: 10.1073/pnas.86.12.4435

Mutations in the maize mitochondrial T-urf13 gene eliminate sensitivity to a fungal pathotoxin.

C J Braun 1, J N Siedow 1, M E Williams 1, C S Levings 3rd 1
PMCID: PMC287284  PMID: 2660137

Abstract

URF13, the product of the mitochondrial T-urf13 gene, confers on Texas cytoplasmic male-steril maize (Zea mays L.) a unique susceptibility to a fungal pathogen (Bipolaris maydis race T) and sensitivity to its pathotoxin. Expression of URF13 in Escherichia coli imparts pathotoxin sensitivity to the bacterium. We show by ion uptake studies in E. coli that a pathotoxin-URF13 interaction causes membrane permeability. Similarly, mitochondrial dysfunction caused by membrane permeabilization probably accounts for increased colonization of maize carrying the Texas cytoplasm by toxin-producing pathogens. Site-directed mutagenesis studies show that approximately one-quarter of the amino acids at the carboxyl end of URF13 can be eliminated without affecting toxin sensitivity. We have identified two dicyclohexylcarbodiimide (DCCD) binding sites in the URF13 protein and show that one of the sites is involved in conferring DCCD protection against the pathotoxin. Substitutional mutations at this DCCD binding site also eliminate toxin sensitivity.

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Selected References

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

  1. Bakker E. P. Membrane potential in a potassium transport-negative mutant of Escherichia coli K-12. The distribution of rubidium in the presence of valinomycin indicates a higher potential than that of the tetraphenylphosphonium cation. Biochim Biophys Acta. 1982 Sep 15;681(3):474–483. doi: 10.1016/0005-2728(82)90190-6. [DOI] [PubMed] [Google Scholar]
  2. Bervillé A., Ghazi A., Charbonnier M., Bonavent J. F. Effects of Methomyl and Helminthosporium maydis Toxin on Matrix Volume, Proton Motive Force, and NAD Accumulation in Maize (Zea mays L.) Mitochondria. Plant Physiol. 1984 Oct;76(2):508–517. doi: 10.1104/pp.76.2.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  4. Dewey R. E., Levings C. S., 3rd, Timothy D. H. Novel recombinations in the maize mitochondrial genome produce a unique transcriptional unit in the Texas male-sterile cytoplasm. Cell. 1986 Feb 14;44(3):439–449. doi: 10.1016/0092-8674(86)90465-4. [DOI] [PubMed] [Google Scholar]
  5. Dewey R. E., Siedow J. N., Timothy D. H., Levings C. S., 3rd A 13-kilodalton maize mitochondrial protein in E. coli confers sensitivity to Bipolaris maydis toxin. Science. 1988 Jan 15;239(4837):293–295. doi: 10.1126/science.3276005. [DOI] [PubMed] [Google Scholar]
  6. Dewey R. E., Timothy D. H., Levings C. S. A mitochondrial protein associated with cytoplasmic male sterility in the T cytoplasm of maize. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5374–5378. doi: 10.1073/pnas.84.15.5374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Engelman D. M., Steitz T. A., Goldman A. Identifying nonpolar transbilayer helices in amino acid sequences of membrane proteins. Annu Rev Biophys Biophys Chem. 1986;15:321–353. doi: 10.1146/annurev.bb.15.060186.001541. [DOI] [PubMed] [Google Scholar]
  8. Fox T. D., Leaver C. J. The Zea mays mitochondrial gene coding cytochrome oxidase subunit II has an intervening sequence and does not contain TGA codons. Cell. 1981 Nov;26(3 Pt 1):315–323. doi: 10.1016/0092-8674(81)90200-2. [DOI] [PubMed] [Google Scholar]
  9. Holden M. J., Sze H. Dissipation of the Membrane Potential in Susceptible Corn Mitochondria by the Toxin of Helminthosporium maydis, Race T, and Toxin Analogs. Plant Physiol. 1987 Jul;84(3):670–676. doi: 10.1104/pp.84.3.670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Holden M. J., Sze H. Helminthosporium maydis T Toxin Increased Membrane Permeability to Ca in Susceptible Corn Mitochondria. Plant Physiol. 1984 May;75(1):235–237. doi: 10.1104/pp.75.1.235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Klein R. R., Koeppe D. E. Mode of Methomyl and Bipolaris maydis (race T) Toxin in Uncoupling Texas Male-Sterile Cytoplasm Corn Mitochondria. Plant Physiol. 1985 Apr;77(4):912–916. doi: 10.1104/pp.77.4.912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Koeppe D. E., Cox J. K., Malone C. P. Mitochondrial heredity: a determinant in the toxic response of maize to the insecticide methomyl. Science. 1978 Sep 29;201(4362):1227–1229. doi: 10.1126/science.201.4362.1227. [DOI] [PubMed] [Google Scholar]
  13. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [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. Laughnan J. R., Gabay-Laughnan S. Cytoplasmic male sterility in maize. Annu Rev Genet. 1983;17:27–48. doi: 10.1146/annurev.ge.17.120183.000331. [DOI] [PubMed] [Google Scholar]
  18. Matthews D. E., Gregory P., Gracen V. E. Helminthosporium maydis Race T Toxin Induces Leakage of NAD from T Cytoplasm Corn Mitochondria. Plant Physiol. 1979 Jun;63(6):1149–1153. doi: 10.1104/pp.63.6.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Miller R. J., Koeppe D. E. Southern corn leaf blight: susceptible and resistant mitochondria. Science. 1971 Jul 2;173(3991):67–69. doi: 10.1126/science.173.3991.67. [DOI] [PubMed] [Google Scholar]
  20. Mohana Rao J. K., Argos P. A conformational preference parameter to predict helices in integral membrane proteins. Biochim Biophys Acta. 1986 Jan 30;869(2):197–214. doi: 10.1016/0167-4838(86)90295-5. [DOI] [PubMed] [Google Scholar]
  21. Nałecz M. J., Casey R. P., Azzi A. Use of N,N'-dicyclohexylcarbodiimide to study membrane-bound enzymes. Methods Enzymol. 1986;125:86–108. doi: 10.1016/s0076-6879(86)25009-0. [DOI] [PubMed] [Google Scholar]
  22. Remaut E., Stanssens P., Fiers W. Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda. Gene. 1981 Oct;15(1):81–93. doi: 10.1016/0378-1119(81)90106-2. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Schneider E., Altendorf K. Proton-conducting portion (F0) from Escherichia coli ATP synthase: preparation, dissociation into subunits, and reconstitution of an active complex. Methods Enzymol. 1986;126:569–578. doi: 10.1016/s0076-6879(86)26059-0. [DOI] [PubMed] [Google Scholar]
  25. Sussman M. R., Strickler J. E., Hager K. M., Slayman C. W. Location of a dicyclohexylcarbodiimide-reactive glutamate residue in the Neurospora crassa plasma membrane H+-ATPase. J Biol Chem. 1987 Apr 5;262(10):4569–4573. [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. Wise R. P., Pring D. R., Gengenbach B. G. Mutation to male fertility and toxin insensitivity in Texas (T)-cytoplasm maize is associated with a frameshift in a mitochondrial open reading frame. Proc Natl Acad Sci U S A. 1987 May;84(9):2858–2862. doi: 10.1073/pnas.84.9.2858. [DOI] [PMC free article] [PubMed] [Google Scholar]

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