Molecular Characterization of Suppressor-Mutator (Spm)-Induced Mutations at the bronze-1 Locus in Maize: The bz-m13 Alleles

  • John W. Schiefelbein
  • Victor Raboy
  • Hwa-Yeong Kim
  • Oliver E. Nelson
Part of the Basic Life Sciences book series (BLSC, volume 47)


The bz-m13 allele of maize contains a defective Suppressor-mutator (dSpm) transposable element and gives rise to a variety of stable and unstable derivatives in the presence of an autonomous Suppressor-mutator (Spm) element. The dSpm-13 element of bz-m13 consists of 2,241 base pairs (bp) and is located within the second exon of the bronze-1 (bz) gene. A number of the stable derivatives, both functional and nonfunctional, derived from bz-m13 were characterized molecularly. Resu ts from genomic DNA blotting experiments indicate that the dSpm-13 element had excised from the locus in each stable derivative analyzed. The unstable derivatives bz-m13CS9 and bz-m13CS6 contain dSpm elements in the same position and orientation as the dSpm-13 element, but they differ in the length of the element. The dSpm-13CS9 element is 902 bp and arose via a deletion between two 5-bp direct repeats within the dSpm-13 element. The dSpm-13CS6 element is 2,239 bp and only differs from dSpm-13 by a 2-bp deletion at the end of one of the 13-bp terminal inverted repeats. The effect of these deletions on the frequency and timing of Spm-induced excision is discussed herein. In the absence of Spm, each of the bz-m13 alleles conditions a nonmutant phenotype despite the presence of the insertions in the second exon. The role of RNA splicing in this phenomenon and the recent finding of an acceptor splice site within the terminal inverted repeat are also discussed.


Transposable Element Terminal Inverted Repeat Acceptor Splice Site Stable Derivative Transposable Element Insertion 
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  1. 1.
    Banks, J., J. Kingsbury, V. Raboy, J.W. Schiefelbein, O. Nelson, Jr., and N. Fedoroff (1985) The Ac and Spm controlling element families in maize. Cold Spring Harbor Symp. Quant. Biol. 50:307–311.PubMedCrossRefGoogle Scholar
  2. 2.
    Brown, J.W.S., G. Feix, and D. Frendewey (1986) Accurate in vitro splicing of two pre-mRNA plant introns in a HeLa cell nuclear extract. EMBO J. 5:2749–2758.PubMedGoogle Scholar
  3. 3.
    Cone, K.C., F.A. Burr, and B. Burr (1986) Molecular analysis of the maize anthocyanin regulatory locus C1. Proc. Natl. Acad. Sci., USA 83:9631–9635.CrossRefGoogle Scholar
  4. 4.
    Dennis, E.S., W.L. Gerlach, W.J. Peacock, and D. Schwartz (1986) Excision of the Ds controlling element from the Adh1 gene of maize. Maydica 31:47–57.Google Scholar
  5. 5.
    Dooner, H.K., and O.E. Nelson (1976) Genetic control of UDPglucose: flavonol 3-O-glucosyltransferase in the endosperm of maize. Biochem. Genet. 15:509–519.CrossRefGoogle Scholar
  6. 6.
    Dooner, H.K., and O.E. Nelson (1979) Heterogeneous flavonoid glucosyltransferases in purple derivatives from a controlling element-suppressed bronze mutant in maize. Proc. Natl. Acad. Sci., USA 76:2369–2371.PubMedCrossRefGoogle Scholar
  7. 7.
    Echt, C.S., and D. Schwartz (1981) Evidence for the inclusion of controlling elements within the structural gene at the waxy locus in maize. Genetics 99:275–284.PubMedGoogle Scholar
  8. 8.
    Fedoroff, N.V., D.B. Furtek, and O.E. Nelson (1984) Cloning of the bronze locus in maize by a simple and generalizable procedure using the transposable controlling element Ac. Proc. Natl. Acad. Sci., USA 81:3825–3829.PubMedCrossRefGoogle Scholar
  9. 9.
    Fedoroff, N., M. Shure, S. Kelly, M. Johns, D. Furtek, J. Schiefelbein, and O. Nelson, Jr. (1984) Isolation of Spm controlling elements from maize. Cold Spring Harbor Symp. Quant. Biol. 49:339–345.PubMedCrossRefGoogle Scholar
  10. 10.
    Frischauf, A., H. Lehrach, A. Poustka, and N. Murray (1983) Lambda replacement vectors carrying polylinker sequences. J. Mol. Biol. 170:827–842.PubMedCrossRefGoogle Scholar
  11. 11.
    Furtek, D. (1986), Cloning and sequence analysis of the bronze locus of maize: Location of transposable element insertions and comparison of two wild type alleles. Ph.D. Thesis, University of Wisconsin, Madison.Google Scholar
  12. 12.
    Gierl, A., Z. Schwarz-Sommer, and H. Saedler (1985) Molecular interactions between the components of the En-I transposable element system of Zea mays. EMBO J. 4:579–583.PubMedGoogle Scholar
  13. 13.
    Henikoff, S. (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359.PubMedCrossRefGoogle Scholar
  14. 14.
    Kim, H.-Y., J.W. Schiefelbein, V. Raboy, D.B. Furtek, and O.E. Nelson (1987) RNA splicing permits expression of a maize gene with a defective Suppressor-mutator transposable element insertion in an exon. Proc. Natl. Acad. Sci., USA 84:5863–5867.PubMedCrossRefGoogle Scholar
  15. 15.
    Klein, A.S., and O.E. Nelson, Jr. (1983) Biochemical consequences of the insertion of a Suppressor-mutator (Spm) receptor at the bronze-1 locus in maize. Proc. Natl. Acad. Sci., USA 80:7591–7595.PubMedCrossRefGoogle Scholar
  16. 16.
    Larson, R.L., and E.H. Coe, Jr. (1977) Gene-dependent flavonoid glucosyl transferase in maize. Biochem. Genet. 15:153–156.PubMedCrossRefGoogle Scholar
  17. 17.
    McClintock, B. (1954) Mutations in maize and chromosomal aberrations in Neurospora. Carnegie Inst. of Washington Yearbook 53:254–260.Google Scholar
  18. 18.
    McClintock, B. (1955) Controlled mutation in maize. Carnegie Inst. of Washington Yearbook 54:245–255.Google Scholar
  19. 19.
    McClintock, B. (1956) Mutation in maize. Carnegie Inst. of Washington Yearbook 55:323–332.Google Scholar
  20. 20.
    McClintock, B. (1961) Further studies of the Suppressor-mutator system of control of gene action in maize. Carnegie Inst. of Washington Yearbook 60:469–476.Google Scholar
  21. 21.
    McClintock, B. (1965) The control of gene action in maize. Brook-haven Symp. Biol. 18:162–184.Google Scholar
  22. 22.
    Nelson, O.E., Jr., and A.S. Klein (1984) Characterization of an Spm-controlled bronze-mutable allele in maize. Genetics 106:769–779.PubMedGoogle Scholar
  23. 23.
    Paz-Ares, J., U. Wienand, P.A. Peterson, and H. Saedler (1986) Molecular cloning of the c locus of Zea mays: A locus regulating the anthocyanin pathway. EMBO J. 5:829–833.PubMedGoogle Scholar
  24. 24.
    Pereira, A., Z. Schwarz-Sommer, A. Gierl, I. Bertram, P.A. Peterson, and H. Saedler (1985) Genetic and molecular analysis of the Enhancer (En) transposable element system of Zea mays. EMBO J. 4:17–23.PubMedGoogle Scholar
  25. 25.
    Pereira, A., H. Cuypers, A. Gierl, Z. Schwarz-Sommer, and H. Saedler (1986) Molecular analysis of the En/Spm transposable element system of Zea mays. EMBO J. 5:835–841.PubMedGoogle Scholar
  26. 26.
    Peterson, P. (1953) A mutable pale green locus in maize. Genetics 38:682–683.Google Scholar
  27. 27.
    Peterson, P. (1960) The pale green mutable system in maize. Genetics 45:115.PubMedGoogle Scholar
  28. 28.
    Peterson, P. (1965) A relationship between the Spm and En control systems in maize. Am. Nat. 99:391–398.CrossRefGoogle Scholar
  29. 29.
    Peterson, P. (1966) Phase variation of regulatory elements in maize. Genetics 54:249–266.PubMedGoogle Scholar
  30. 30.
    Saedler, H., and P. Nevers (1985) Transposition in plants: A molecular model. EMBO J. 4:585–590.PubMedGoogle Scholar
  31. 31.
    Sanger, F., S. Nicklen, and A.R. Coulson (1977) DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci., USA 74:5463–5467.PubMedCrossRefGoogle Scholar
  32. 32.
    Schiefelbein, J.W., V. Raboy, N.V. Fedoroff, and O.E. Nelson, Jr. (1985) Deletions within a defective Suppressor-mutator element in maize affect the frequency and developmental timing of its excision from the bronze locus. Proc. Natl. Acad. Sci., USA 82:4783–4787.PubMedCrossRefGoogle Scholar
  33. 33.
    Schiefelbein, J.W., D.B. Furtek, V. Raboy, J.A. Banks, N.V. Fedoroff, and O.E. Nelson (1985) Exploiting transposable elements to study the expression of a maize gene. In Plant Genetics, M. Freeling, ed. Alan R. Liss, Inc., New York, pp. 445–459.Google Scholar
  34. 34.
    Schwarz-Sommer, Z., A. Gierl, R.B. Klösgen, U. Wienand, P.A. Peterson, and H. Saedler (1984) The Spm (En) transposable element controls the excision of a 2-kb DNA insert at the wxm8 allele of Zea mays. EMBO J. 3:1021–1028.PubMedGoogle Scholar
  35. 35.
    Schwarz-Sommer, Z., A. Gierl, H. Cuypers, P.A. Peterson, and H. Saedler (1985) Plant transposable elements generate the DNA sequence diversity needed in evolution. EMBO J. 4:591–597.PubMedGoogle Scholar
  36. 36.
    Schwarz-Sommer, Z., A. Gierl, R. Berndtgen, and H. Saedler (1985) Sequence comparison of “states” of a1-m1 suggests a model of Spm (En) action. EMBO J. 4:2439–2443.PubMedGoogle Scholar
  37. 37.
    Schwarz-Sommer, Z., N. Shepherd, E. Tacke, A. Gierl, W. Rohde, L. Leclercq, M. Mattes, R. Berndtgen, P.A. Peterson, and H. Saedler (1987) Influence of transposable elements on the structure and function of the A1 gene of Zea mays. EMBO J. 6:287–294.PubMedGoogle Scholar
  38. 38.
    Shapiro, J.A., ed. (1983) Mobile Genetic Elements. Academic Press, New York.Google Scholar
  39. 39.
    Tacke, E., Z. Schwarz-Sommer, P.A. Peterson, and H. Saedler (1986) Molecular analysis of states of the A1 locus of Zea mays. Maydica 31:83–91.Google Scholar
  40. 40.
    Tuschall, D.M., and L.C. Hannah (1982) Altered maize endosperm ADP-glucose pyrophosphorylases from revertants of a Shrunken-2-dissociation allele. Genetics 100:105–111.PubMedGoogle Scholar
  41. 41.
    Vieira, J., and J. Messing (1982) The pUC plasmids, and M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268.PubMedCrossRefGoogle Scholar
  42. 42.
    Wessler, S.R., G. Baran, M. Varagona, and S.L. Dellaporta (1986) Excision of DS produces waxy proteins with a range of enzymatic activities. EMBO J. 5:2427–2432.PubMedGoogle Scholar
  43. 43.
    Wessler, S.R., G. Baran, and M. Varagona (1987) The maize trans-posable element DS is spliced from RNA. Science 237:916–918.PubMedCrossRefGoogle Scholar
  44. 44.
    Yanisch-Perron, C., J. Vieira, and J. Messing (1985) Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • John W. Schiefelbein
    • 1
  • Victor Raboy
    • 1
  • Hwa-Yeong Kim
    • 1
  • Oliver E. Nelson
    • 1
  1. 1.Laboratory of GeneticsUniversity of WisconsinMadisonUSA

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