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Plant Cell. Apr 1997; 9(4): 611–625.
PMCID: PMC156943

Evidence for direct activation of an anthocyanin promoter by the maize C1 protein and comparison of DNA binding by related Myb domain proteins.


The enzyme-encoding genes of two classes of maize flavonoid pigments, anthocyanins and phlobaphenes, are differentially regulated by distinct transcription factors. Anthocyanin biosynthetic gene activation requires the Myb domain C1 protein and the basic helix-loop-helix B or R proteins. In the phlobaphene pathway, a subset of C1-regulated genes, including a1, are activated by the Myb domain P protein independently of B/R. We show sequence-specific binding to the a1 promoter by C1 in the absence of B. Activation is decreased by mutations in the C1 DNA binding domain or in a1 sequences bound by C1, providing direct evidence for activation of the anthocyanin biosynthetic genes by C1. The two C1 binding sites in the a1 promoter are also bound by P. One site is bound with higher affinity by P relative to C1, whereas the other site is bound with similar lower affinity by both proteins. Interestingly, either site is sufficient for C1 plus B/R or P activation in vivo, demonstrating that differences in DNA binding affinities between P and C1 are insufficient to explain the differential requirement for B. Results of DNA binding site-selection experiments suggest that C1 has a broader DNA binding specificity than does P, which may help C1 to activate a more diverse set of promoters.

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Selected References

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  • Abate C, Luk D, Gentz R, Rauscher FJ, 3rd, Curran T. Expression and purification of the leucine zipper and DNA-binding domains of Fos and Jun: both Fos and Jun contact DNA directly. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1032–1036. [PMC free article] [PubMed]
  • Ames P, Parkinson JS. Constitutively signaling fragments of Tsr, the Escherichia coli serine chemoreceptor. J Bacteriol. 1994 Oct;176(20):6340–6348. [PMC free article] [PubMed]
  • Biedenkapp H, Borgmeyer U, Sippel AE, Klempnauer KH. Viral myb oncogene encodes a sequence-specific DNA-binding activity. Nature. 1988 Oct 27;335(6193):835–837. [PubMed]
  • Bodeau JP, Walbot V. Regulated transcription of the maize Bronze-2 promoter in electroporated protoplasts requires the C1 and R gene products. Mol Gen Genet. 1992 Jun;233(3):379–387. [PubMed]
  • Callis J, Fromm M, Walbot V. Introns increase gene expression in cultured maize cells. Genes Dev. 1987 Dec;1(10):1183–1200. [PubMed]
  • Chandler VL, Radicella JP, Robbins TP, 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. [PMC free article] [PubMed]
  • Cone KC, Cocciolone SM, Burr FA, Burr B. Maize anthocyanin regulatory gene pl is a duplicate of c1 that functions in the plant. Plant Cell. 1993 Dec;5(12):1795–1805. [PMC free article] [PubMed]
  • Dooner HK, Robbins TP, Jorgensen RA. Genetic and developmental control of anthocyanin biosynthesis. Annu Rev Genet. 1991;25:173–199. [PubMed]
  • Garcia A, LaMontagne K, Reavis D, Stober-Grässer U, Lipsick JS. Determinants of sequence-specific DNA-binding by p48v-myb. Oncogene. 1991 Feb;6(2):265–273. [PubMed]
  • Gill G, Ptashne M. Mutants of GAL4 protein altered in an activation function. Cell. 1987 Oct 9;51(1):121–126. [PubMed]
  • Goff SA, Klein TM, Roth BA, Fromm ME, Cone KC, Radicella JP, Chandler VL. Transactivation of anthocyanin biosynthetic genes following transfer of B regulatory genes into maize tissues. EMBO J. 1990 Aug;9(8):2517–2522. [PMC free article] [PubMed]
  • Goff SA, Cone KC, Chandler VL. 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. [PubMed]
  • Grotewold E, Athma P, Peterson T. Alternatively spliced products of the maize P gene encode proteins with homology to the DNA-binding domain of myb-like transcription factors. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4587–4591. [PMC free article] [PubMed]
  • Grotewold E, Drummond BJ, Bowen B, Peterson T. The myb-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonoid biosynthetic gene subset. Cell. 1994 Feb 11;76(3):543–553. [PubMed]
  • Hoopes BC, LeBlanc JF, Hawley DK. Kinetic analysis of yeast TFIID-TATA box complex formation suggests a multi-step pathway. J Biol Chem. 1992 Jun 5;267(16):11539–11547. [PubMed]
  • Howe KM, Reakes CF, Watson RJ. Characterization of the sequence-specific interaction of mouse c-myb protein with DNA. EMBO J. 1990 Jan;9(1):161–169. [PMC free article] [PubMed]
  • Klein TM, Roth BA, Fromm ME. Regulation of anthocyanin biosynthetic genes introduced into intact maize tissues by microprojectiles. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6681–6685. [PMC free article] [PubMed]
  • Loake GJ, Faktor O, Lamb CJ, Dixon RA. Combination of H-box [CCTACC(N)7CT] and G-box (CACGTG) cis elements is necessary for feed-forward stimulation of a chalcone synthase promoter by the phenylpropanoid-pathway intermediate p-coumaric acid. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9230–9234. [PMC free article] [PubMed]
  • Lois R, Dietrich A, Hahlbrock K, Schulz W. A phenylalanine ammonia-lyase gene from parsley: structure, regulation and identification of elicitor and light responsive cis-acting elements. EMBO J. 1989 Jun;8(6):1641–1648. [PMC free article] [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Ludwig SR, Wessler SR. Maize R gene family: tissue-specific helix-loop-helix proteins. Cell. 1990 Sep 7;62(5):849–851. [PubMed]
  • Ludwig SR, Habera LF, Dellaporta SL, Wessler SR. 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. [PMC free article] [PubMed]
  • Luehrsen KR, de Wet JR, Walbot V. Transient expression analysis in plants using firefly luciferase reporter gene. Methods Enzymol. 1992;216:397–414. [PubMed]
  • Ogata K, Morikawa S, Nakamura H, Sekikawa A, Inoue T, Kanai H, Sarai A, Ishii S, Nishimura Y. Solution structure of a specific DNA complex of the Myb DNA-binding domain with cooperative recognition helices. Cell. 1994 Nov 18;79(4):639–648. [PubMed]
  • Patterson GI, Thorpe CJ, Chandler VL. Paramutation, an allelic interaction, is associated with a stable and heritable reduction of transcription of the maize b regulatory gene. Genetics. 1993 Nov;135(3):881–894. [PMC free article] [PubMed]
  • Paz-Ares J, Ghosal D, Saedler H. Molecular analysis of the C1-I allele from Zea mays: a dominant mutant of the regulatory C1 locus. EMBO J. 1990 Feb;9(2):315–321. [PMC free article] [PubMed]
  • Pollock R, Treisman R. A sensitive method for the determination of protein-DNA binding specificities. Nucleic Acids Res. 1990 Nov 11;18(21):6197–6204. [PMC free article] [PubMed]
  • Roth BA, Goff SA, Klein TM, Fromm ME. C1- and R-dependent expression of the maize Bz1 gene requires sequences with homology to mammalian myb and myc binding sites. Plant Cell. 1991 Mar;3(3):317–325. [PMC free article] [PubMed]
  • Sablowski RW, Moyano E, Culianez-Macia FA, Schuch W, Martin C, Bevan M. A flower-specific Myb protein activates transcription of phenylpropanoid biosynthetic genes. EMBO J. 1994 Jan 1;13(1):128–137. [PMC free article] [PubMed]
  • Sainz MB, Goff SA, Chandler VL. Extensive mutagenesis of a transcriptional activation domain identifies single hydrophobic and acidic amino acids important for activation in vivo. Mol Cell Biol. 1997 Jan;17(1):115–122. [PMC free article] [PubMed]
  • Schwarz-Sommer Z, Shepherd N, Tacke E, Gierl A, Rohde W, Leclercq L, Mattes M, Berndtgen R, Peterson PA, Saedler H. Influence of transposable elements on the structure and function of the A1 gene of Zea mays. EMBO J. 1987 Feb;6(2):287–294. [PMC free article] [PubMed]
  • Shieh MW, Wessler SR, Raikhel NV. Nuclear targeting of the maize R protein requires two nuclear localization sequences. Plant Physiol. 1993 Feb;101(2):353–361. [PMC free article] [PubMed]
  • Tuerck JA, Fromm ME. Elements of the maize A1 promoter required for transactivation by the anthocyanin B/C1 or phlobaphene P regulatory genes. Plant Cell. 1994 Nov;6(11):1655–1663. [PMC free article] [PubMed]
  • Vieira J, Messing J. New pUC-derived cloning vectors with different selectable markers and DNA replication origins. Gene. 1991 Apr;100:189–194. [PubMed]
  • Weintraub H, Davis R, Tapscott S, Thayer M, Krause M, Benezra R, Blackwell TK, Turner D, Rupp R, Hollenberg S, et al. The myoD gene family: nodal point during specification of the muscle cell lineage. Science. 1991 Feb 15;251(4995):761–766. [PubMed]
  • Williams CE, Grotewold E. Differences between plant and animal Myb domains are fundamental for DNA binding activity, and chimeric Myb domains have novel DNA binding specificities. J Biol Chem. 1997 Jan 3;272(1):563–571. [PubMed]

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