Abstract
The recessive mutation intensifier1 of maize apparently causes an overall increase in flavonoid production in the aleurone. The mechanism by which this is achieved is not understood. We have succeeded in cloning the intensifier1 gene by transposon tagging with Suppressor-mutator and found, by sequence analyses, that it shares homology with known transcription factors in the anthocyanin pathway, in particular the r1/b1 multigene family in maize. Two cDNAs and a genomic clone were completely sequenced, and together they showed that the transcripts were misspliced. The frequency of missplicing was investigated by polymerase chain reaction analyses and sequencing of the individual introns. These studies indicate that very little functional transcript was made. Indeed, missplicing may be a mechanism for reducing the levels of a transcription factor that, when present, acts as a repressor of anthocyanin biosynthesis.
Full Text
The Full Text of this article is available as a PDF (2.3 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
- Benezra R., Davis R. L., Lockshon D., Turner D. L., Weintraub H. The protein Id: a negative regulator of helix-loop-helix DNA binding proteins. Cell. 1990 Apr 6;61(1):49–59. doi: 10.1016/0092-8674(90)90214-y. [DOI] [PubMed] [Google Scholar]
- Bingham P. M., Chou T. B., Mims I., Zachar Z. On/off regulation of gene expression at the level of splicing. Trends Genet. 1988 May;4(5):134–138. doi: 10.1016/0168-9525(88)90136-9. [DOI] [PubMed] [Google Scholar]
- Burr B., Burr F. A. Controlling-element events at the shrunken locus in maize. Genetics. 1981 May;98(1):143–156. doi: 10.1093/genetics/98.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burr B., Burr F. A. Recombinant inbreds for molecular mapping in maize: theoretical and practical considerations. Trends Genet. 1991 Feb;7(2):55–60. doi: 10.1016/0168-9525(91)90232-F. [DOI] [PubMed] [Google Scholar]
- Burr B., Burr F. A., Thompson K. H., Albertson M. C., Stuber C. W. Gene mapping with recombinant inbreds in maize. Genetics. 1988 Mar;118(3):519–526. doi: 10.1093/genetics/118.3.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chandler V. L., Radicella J. P., Robbins T. P., Chen J., Turks D. Two regulatory genes of the maize anthocyanin pathway are homologous: isolation of B utilizing R genomic sequences. Plant Cell. 1989 Dec;1(12):1175–1183. doi: 10.1105/tpc.1.12.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chou T. B., Zachar Z., Bingham P. M. Developmental expression of a regulatory gene is programmed at the level of splicing. EMBO J. 1987 Dec 20;6(13):4095–4104. doi: 10.1002/j.1460-2075.1987.tb02755.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Consonni G., Geuna F., Gavazzi G., Tonelli C. Molecular homology among members of the R gene family in maize. Plant J. 1993 Feb;3(2):335–346. doi: 10.1111/j.1365-313x.1993.tb00185.x. [DOI] [PubMed] [Google Scholar]
- Consonni G., Viotti A., Dellaporta S. L., Tonelli C. cDNA nucleotide sequence of Sn, a regulatory gene in maize. Nucleic Acids Res. 1992 Jan 25;20(2):373–373. doi: 10.1093/nar/20.2.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dooner H. K., Nelson O. E. Genetic control of UDPglucose:flavonol 3-O-glucosyltransferase in the endosperm of maize. Biochem Genet. 1977 Jun;15(5-6):509–519. doi: 10.1007/BF00520194. [DOI] [PubMed] [Google Scholar]
- Goff S. A., Cone K. C., Chandler V. L. Functional analysis of the transcriptional activator encoded by the maize B gene: evidence for a direct functional interaction between two classes of regulatory proteins. Genes Dev. 1992 May;6(5):864–875. doi: 10.1101/gad.6.5.864. [DOI] [PubMed] [Google Scholar]
- Goodall G. J., Filipowicz W. Different effects of intron nucleotide composition and secondary structure on pre-mRNA splicing in monocot and dicot plants. EMBO J. 1991 Sep;10(9):2635–2644. doi: 10.1002/j.1460-2075.1991.tb07806.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goodall G. J., Filipowicz W. The AU-rich sequences present in the introns of plant nuclear pre-mRNAs are required for splicing. Cell. 1989 Aug 11;58(3):473–483. doi: 10.1016/0092-8674(89)90428-5. [DOI] [PubMed] [Google Scholar]
- Goodrich J., Carpenter R., Coen E. S. A common gene regulates pigmentation pattern in diverse plant species. Cell. 1992 Mar 6;68(5):955–964. doi: 10.1016/0092-8674(92)90038-e. [DOI] [PubMed] [Google Scholar]
- Hanley B. A., Schuler M. A. Plant intron sequences: evidence for distinct groups of introns. Nucleic Acids Res. 1988 Jul 25;16(14B):7159–7176. doi: 10.1093/nar/16.14.7159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kim J. B., Spotts G. D., Halvorsen Y. D., Shih H. M., Ellenberger T., Towle H. C., Spiegelman B. M. Dual DNA binding specificity of ADD1/SREBP1 controlled by a single amino acid in the basic helix-loop-helix domain. Mol Cell Biol. 1995 May;15(5):2582–2588. doi: 10.1128/mcb.15.5.2582. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kozak M. At least six nucleotides preceding the AUG initiator codon enhance translation in mammalian cells. J Mol Biol. 1987 Aug 20;196(4):947–950. doi: 10.1016/0022-2836(87)90418-9. [DOI] [PubMed] [Google Scholar]
- Larson R. L., Coe E. H., Jr Gene-dependent flavonoid glucosyltransferase in maize. Biochem Genet. 1977 Feb;15(1-2):153–156. doi: 10.1007/BF00484558. [DOI] [PubMed] [Google Scholar]
- Larson R., Bussard J. B., Coe E. H., Jr Gene-dependent flavonoid 3'-hydroxylation in maize. Biochem Genet. 1986 Aug;24(7-8):615–624. doi: 10.1007/BF00504338. [DOI] [PubMed] [Google Scholar]
- Ludwig S. R., Habera L. F., Dellaporta S. L., Wessler S. R. Lc, a member of the maize R gene family responsible for tissue-specific anthocyanin production, encodes a protein similar to transcriptional activators and contains the myc-homology region. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7092–7096. doi: 10.1073/pnas.86.18.7092. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Luehrsen K. R., Walbot V. Addition of A- and U-rich sequence increases the splicing efficiency of a deleted form of a maize intron. Plant Mol Biol. 1994 Feb;24(3):449–463. doi: 10.1007/BF00024113. [DOI] [PubMed] [Google Scholar]
- Luehrsen K. R., Walbot V. Intron creation and polyadenylation in maize are directed by AU-rich RNA. Genes Dev. 1994 May 1;8(9):1117–1130. doi: 10.1101/gad.8.9.1117. [DOI] [PubMed] [Google Scholar]
- Oellers N., Dehio M., Knust E. bHLH proteins encoded by the Enhancer of split complex of Drosophila negatively interfere with transcriptional activation mediated by proneural genes. Mol Gen Genet. 1994 Sep 1;244(5):465–473. doi: 10.1007/BF00583897. [DOI] [PubMed] [Google Scholar]
- Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perrot G. H., Cone K. C. Nucleotide sequence of the maize R-S gene. Nucleic Acids Res. 1989 Oct 11;17(19):8003–8003. doi: 10.1093/nar/17.19.8003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Radicella J. P., Turks D., Chandler V. L. Cloning and nucleotide sequence of a cDNA encoding B-Peru, a regulatory protein of the anthocyanin pathway in maize. Plant Mol Biol. 1991 Jul;17(1):127–130. doi: 10.1007/BF00036813. [DOI] [PubMed] [Google Scholar]
- Schmidt R. J., Burr F. A., Aukerman M. J., Burr B. Maize regulatory gene opaque-2 encodes a protein with a "leucine-zipper" motif that binds to zein DNA. Proc Natl Acad Sci U S A. 1990 Jan;87(1):46–50. doi: 10.1073/pnas.87.1.46. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Sutherland J. C., Lin B., Monteleone D. C., Mugavero J., Sutherland B. M., Trunk J. Electronic imaging system for direct and rapid quantitation of fluorescence from electrophoretic gels: application to ethidium bromide-stained DNA. Anal Biochem. 1987 Jun;163(2):446–457. doi: 10.1016/0003-2697(87)90247-8. [DOI] [PubMed] [Google Scholar]