Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1990 Feb;9(2):315–321. doi: 10.1002/j.1460-2075.1990.tb08113.x

Molecular analysis of the C1-I allele from Zea mays: a dominant mutant of the regulatory C1 locus.

J Paz-Ares 1, D Ghosal 1, H Saedler 1
PMCID: PMC551668  PMID: 2303027

Abstract

The C1 locus of Zea mays (maize) controls the expression of genes involved in anthocyanin biosynthesis in aleurone and scutellar tissue and encodes a protein with the features of a transcriptional activator. C1-I is a dominant negative mutant which inhibits pigment formation. The structure of the C1-I allele was determined by cloning and sequencing of this allele and of two distinct C1-I derived cDNAs. C1-I has two major and several minor sequence differences with respect to the wild-type C1 allele. Transcription initiation occurs at the same position as in wild-type but transcription yields two different products, one major RNA of 1.3 kb and one minor RNA of 1.45 kb in length, encoding two proteins of 252 and 108 amino acids respectively. The longer 252 amino acid C1-I protein differs from the 273 amino acid wild-type C1 protein at several positions but most prominently at its carboxy terminus, resulting in reduced acidity of the C1-I protein. A similar change in acidity of the Gal4 protein of yeast converted this transcriptional activator into a repressor protein. We discuss the dominant phenotype of C1-I with respect to its possible repressor function in contrast to the activator function of the C1 gene product.

Full text

PDF
315

Images in this article

316Fig. 1.
on p.316
318Fig. 3.
on p.318
318Fig. 4.
on p.318
319Fig. 5.
on p.319

Selected References

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

  1. Biedenkapp H., Borgmeyer U., Sippel A. E., Klempnauer K. H. Viral myb oncogene encodes a sequence-specific DNA-binding activity. Nature. 1988 Oct 27;335(6193):835–837. doi: 10.1038/335835a0. [DOI] [PubMed] [Google Scholar]
  2. Coe E H. Spontaneous Mutation of the Aleurone Color Inhibitor in Maize. Genetics. 1962 Jun;47(6):779–783. doi: 10.1093/genetics/47.6.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cone K. C., Burr F. A., Burr B. Molecular analysis of the maize anthocyanin regulatory locus C1. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9631–9635. doi: 10.1073/pnas.83.24.9631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Courey A. J., Tjian R. Analysis of Sp1 in vivo reveals multiple transcriptional domains, including a novel glutamine-rich activation motif. Cell. 1988 Dec 2;55(5):887–898. doi: 10.1016/0092-8674(88)90144-4. [DOI] [PubMed] [Google Scholar]
  5. Davis R. W., Hyman R. W. A study in evolution: the DNA base sequence homology between coliphages T7 and T3. J Mol Biol. 1971 Dec 14;62(2):287–301. doi: 10.1016/0022-2836(71)90428-1. [DOI] [PubMed] [Google Scholar]
  6. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dooner H. K., Nelson O. E. Interaction among C, R and Vp in the Control of the Bz Glucosyltransferase during Endosperm Development in Maize. Genetics. 1979 Feb;91(2):309–315. doi: 10.1093/genetics/91.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eisenberg D. Three-dimensional structure of membrane and surface proteins. Annu Rev Biochem. 1984;53:595–623. doi: 10.1146/annurev.bi.53.070184.003115. [DOI] [PubMed] [Google Scholar]
  9. Fedoroff N. V., Furtek D. B., Nelson O. E. Cloning of the bronze locus in maize by a simple and generalizable procedure using the transposable controlling element Activator (Ac). Proc Natl Acad Sci U S A. 1984 Jun;81(12):3825–3829. doi: 10.1073/pnas.81.12.3825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  11. Frischauf A. M., Lehrach H., Poustka A., Murray N. Lambda replacement vectors carrying polylinker sequences. J Mol Biol. 1983 Nov 15;170(4):827–842. doi: 10.1016/s0022-2836(83)80190-9. [DOI] [PubMed] [Google Scholar]
  12. Garnier J., Osguthorpe D. J., Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. doi: 10.1016/0022-2836(78)90297-8. [DOI] [PubMed] [Google Scholar]
  13. Giniger E., Ptashne M. Transcription in yeast activated by a putative amphipathic alpha helix linked to a DNA binding unit. Nature. 1987 Dec 17;330(6149):670–672. doi: 10.1038/330670a0. [DOI] [PubMed] [Google Scholar]
  14. Gribskov M., Devereux J., Burgess R. R. The codon preference plot: graphic analysis of protein coding sequences and prediction of gene expression. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):539–549. doi: 10.1093/nar/12.1part2.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Hope I. A., Mahadevan S., Struhl K. Structural and functional characterization of the short acidic transcriptional activation region of yeast GCN4 protein. Nature. 1988 Jun 16;333(6174):635–640. doi: 10.1038/333635a0. [DOI] [PubMed] [Google Scholar]
  17. Keegan L., Gill G., Ptashne M. Separation of DNA binding from the transcription-activating function of a eukaryotic regulatory protein. Science. 1986 Feb 14;231(4739):699–704. doi: 10.1126/science.3080805. [DOI] [PubMed] [Google Scholar]
  18. Klempnauer K. H., Sippel A. E. The highly conserved amino-terminal region of the protein encoded by the v-myb oncogene functions as a DNA-binding domain. EMBO J. 1987 Sep;6(9):2719–2725. doi: 10.1002/j.1460-2075.1987.tb02565.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  20. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  21. O'Reilly C., Shepherd N. S., Pereira A., Schwarz-Sommer Z., Bertram I., Robertson D. S., Peterson P. A., Saedler H. Molecular cloning of the a1 locus of Zea mays using the transposable elements En and Mu1. EMBO J. 1985 Apr;4(4):877–882. doi: 10.1002/j.1460-2075.1985.tb03713.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Paz-Ares J., Ghosal D., Wienand U., Peterson P. A., Saedler H. The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators. EMBO J. 1987 Dec 1;6(12):3553–3558. doi: 10.1002/j.1460-2075.1987.tb02684.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Paz-Ares J., Wienand U., Peterson P. A., Saedler H. Molecular cloning of the c locus of Zea mays: a locus regulating the anthocyanin pathway. EMBO J. 1986 May;5(5):829–833. doi: 10.1002/j.1460-2075.1986.tb04291.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ptashne M. How eukaryotic transcriptional activators work. Nature. 1988 Oct 20;335(6192):683–689. doi: 10.1038/335683a0. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Schwarz-Sommer Z., Gierl A., Cuypers H., Peterson P. A., Saedler H. Plant transposable elements generate the DNA sequence diversity needed in evolution. EMBO J. 1985 Mar;4(3):591–597. doi: 10.1002/j.1460-2075.1985.tb03671.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schwarz-Sommer Z., Gierl A., Klösgen R. B., Wienand U., Peterson P. A., Saedler H. The Spm (En) transposable element controls the excision of a 2-kb DNA insert at the wx allele of Zea mays. EMBO J. 1984 May;3(5):1021–1028. doi: 10.1002/j.1460-2075.1984.tb01922.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Struhl K. Promoters, activator proteins, and the mechanism of transcriptional initiation in yeast. Cell. 1987 May 8;49(3):295–297. doi: 10.1016/0092-8674(87)90277-7. [DOI] [PubMed] [Google Scholar]
  29. Struhl K. The DNA-binding domains of the jun oncoprotein and the yeast GCN4 transcriptional activator protein are functionally homologous. Cell. 1987 Sep 11;50(6):841–846. doi: 10.1016/0092-8674(87)90511-3. [DOI] [PubMed] [Google Scholar]
  30. Theres N, Scheele T, Starlinger P. Cloning of the Bz2 locus of Zea mays using the transposable element Ds as a gene tag. Mol Gen Genet. 1987 Aug;209(1):193–197. doi: 10.1007/BF00329858. [DOI] [PubMed] [Google Scholar]
  31. Vogt P. K., Bos T. J., Doolittle R. F. Homology between the DNA-binding domain of the GCN4 regulatory protein of yeast and the carboxyl-terminal region of a protein coded for by the oncogene jun. Proc Natl Acad Sci U S A. 1987 May;84(10):3316–3319. doi: 10.1073/pnas.84.10.3316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES