THE PLANT CELL, Vol 9, Issue 4 611-625, Copyright © 1997 by American Society of Plant Biologists

RESEARCH ARTICLE

Evidence for Direct Activation of an Anthocyanin Promoter by the Maize C1 Protein and Comparison of DNA Binding by Related Myb Domain Proteins

M. B. Sainz, E. Grotewold and V. L. Chandler
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403

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.

This article has been cited by other articles:

				H. Ma, M. Pooler, and R. Griesbach Ratio of Myc and Myb Transcription Factors Regulates Anthocyanin Production in Orchid Flowers J. Amer. Soc. Hort. Sci., January 1, 2008; 133(1): 133 - 138. [Abstract] [Full Text] [PDF]

				J. M. Hernandez, A. Feller, K. Morohashi, K. Frame, and E. Grotewold The basic helix loop helix domain of maize R links transcriptional regulation and histone modifications by recruitment of an EMSY-related factor PNAS, October 23, 2007; 104(43): 17222 - 17227. [Abstract] [Full Text] [PDF]

				S. Shimada, H. Otsuki, and M. Sakuta Transcriptional control of anthocyanin biosynthetic genes in the Caryophyllales J. Exp. Bot., March 1, 2007; 58(5): 957 - 967. [Abstract] [Full Text] [PDF]

				H. Mano, F. Ogasawara, K. Sato, H. Higo, and Y. Minobe Isolation of a Regulatory Gene of Anthocyanin Biosynthesis in Tuberous Roots of Purple-Fleshed Sweet Potato Plant Physiology, March 1, 2007; 143(3): 1252 - 1268. [Abstract] [Full Text] [PDF]

				A. Feller, J. M. Hernandez, and E. Grotewold An ACT-like Domain Participates in the Dimerization of Several Plant Basic-helix-loop-helix Transcription Factors J. Biol. Chem., September 29, 2006; 281(39): 28964 - 28974. [Abstract] [Full Text] [PDF]

				F. Quattrocchio, W. Verweij, A. Kroon, C. Spelt, J. Mol, and R. Koes PH4 of Petunia Is an R2R3 MYB Protein That Activates Vacuolar Acidification through Interactions with Basic-Helix-Loop-Helix Transcription Factors of the Anthocyanin Pathway PLANT CELL, May 1, 2006; 18(5): 1274 - 1291. [Abstract] [Full Text] [PDF]

				K. Schwinn, J. Venail, Y. Shang, S. Mackay, V. Alm, E. Butelli, R. Oyama, P. Bailey, K. Davies, and C. Martin A Small Family of MYB-Regulatory Genes Controls Floral Pigmentation Intensity and Patterning in the Genus Antirrhinum PLANT CELL, April 1, 2006; 18(4): 831 - 851. [Abstract] [Full Text] [PDF]

				S.-J. Ong, H.-M. Hsu, H.-W. Liu, C.-H. Chu, and J.-H. Tai Multifarious Transcriptional Regulation of Adhesion Protein Gene ap65-1 by a Novel Myb1 Protein in the Protozoan Parasite Trichomonas vaginalis Eukaryot. Cell, February 1, 2006; 5(2): 391 - 399. [Abstract] [Full Text] [PDF]

				K. H. Ryu, Y. H. Kang, Y.-h. Park, I. Hwang, J. Schiefelbein, and M. M. Lee The WEREWOLF MYB protein directly regulates CAPRICE transcription during cell fate specification in the Arabidopsis root epidermis Development, November 1, 2005; 132(21): 4765 - 4775. [Abstract] [Full Text] [PDF]

				N. O. Steffens, C. Galuschka, M. Schindler, L. Bulow, and R. Hehl AthaMap web tools for database-assisted identification of combinatorial cis-regulatory elements and the display of highly conserved transcription factor binding sites in Arabidopsis thaliana Nucleic Acids Res., July 1, 2005; 33(suppl_2): W397 - W402. [Abstract] [Full Text] [PDF]

				Y. Koshino-Kimura, T. Wada, T. Tachibana, R. Tsugeki, S. Ishiguro, and K. Okada Regulation of CAPRICE Transcription by MYB Proteins for Root Epidermis Differentiation in Arabidopsis Plant Cell Physiol., June 1, 2005; 46(6): 817 - 826. [Abstract] [Full Text] [PDF]

				J. C. Verdonk, M. A. Haring, A. J. van Tunen, and R. C. Schuurink ODORANT1 Regulates Fragrance Biosynthesis in Petunia Flowers PLANT CELL, May 1, 2005; 17(5): 1612 - 1624. [Abstract] [Full Text] [PDF]

				Z. Swigonova, J. L. Bennetzen, and J. Messing Structure and Evolution of the r/b Chromosomal Regions in Rice, Maize and Sorghum Genetics, February 1, 2005; 169(2): 891 - 906. [Abstract] [Full Text] [PDF]

				J. M. Hernandez, G. F. Heine, N. G. Irani, A. Feller, M.-G. Kim, T. Matulnik, V. L. Chandler, and E. Grotewold Different Mechanisms Participate in the R-dependent Activity of the R2R3 MYB Transcription Factor C1 J. Biol. Chem., November 12, 2004; 279(46): 48205 - 48213. [Abstract] [Full Text] [PDF]

				G. F. Heine, J. M. Hernandez, and E. Grotewold Two Cysteines in Plant R2R3 MYB Domains Participate in REDOX-dependent DNA Binding J. Biol. Chem., September 3, 2004; 279(36): 37878 - 37885. [Abstract] [Full Text] [PDF]

				C. D. Goodman, P. Casati, and V. Walbot A Multidrug Resistance-Associated Protein Involved in Anthocyanin Transport in Zea mays PLANT CELL, July 1, 2004; 16(7): 1812 - 1826. [Abstract] [Full Text] [PDF]

				E. Himi and K. Noda Isolation and location of three homoeologous dihydroflavonol-4-reductase (DFR) genes of wheat and their tissue-dependent expression J. Exp. Bot., February 1, 2004; 55(396): 365 - 375. [Abstract] [Full Text] [PDF]

				C. C. Carey, J. T. Strahle, D. A. Selinger, and V. L. Chandler Mutations in the pale aleurone color1 Regulatory Gene of the Zea mays Anthocyanin Pathway Have Distinct Phenotypes Relative to the Functionally Similar TRANSPARENT TESTA GLABRA1 Gene in Arabidopsis thaliana PLANT CELL, February 1, 2004; 16(2): 450 - 464. [Abstract] [Full Text] [PDF]

				P. Elomaa, A. Uimari, M. Mehto, V. A. Albert, R. A.E. Laitinen, and T. H. Teeri Activation of Anthocyanin Biosynthesis in Gerbera hybrida (Asteraceae) Suggests Conserved Protein-Protein and Protein-Promoter Interactions between the Anciently Diverged Monocots and Eudicots Plant Physiology, December 1, 2003; 133(4): 1831 - 1842. [Abstract] [Full Text]

				H. Guo and S. P. Moose Conserved Noncoding Sequences among Cultivated Cereal Genomes Identify Candidate Regulatory Sequence Elements and Patterns of Promoter Evolution PLANT CELL, May 1, 2003; 15(5): 1143 - 1158. [Abstract] [Full Text]

				M. A. Heim, M. Jakoby, M. Werber, C. Martin, B. Weisshaar, and P. C. Bailey The Basic Helix-Loop-Helix Transcription Factor Family in Plants: A Genome-Wide Study of Protein Structure and Functional Diversity Mol. Biol. Evol., May 1, 2003; 20(5): 735 - 747. [Abstract] [Full Text] [PDF]

				W. Pooma, C. Gersos, and E. Grotewold Transposon Insertions in the Promoter of the Zea mays a1 Gene Differentially Affect Transcription by the Myb Factors P and C1 Genetics, June 1, 2002; 161(2): 793 - 801. [Abstract] [Full Text] [PDF]

				T. Wada, T. Kurata, R. Tominaga, Y. Koshino-Kimura, T. Tachibana, K. Goto, M. D. Marks, Y. Shimura, and K. Okada Role of a positive regulator of root hair development, CAPRICE, in Arabidopsis root epidermal cell differentiation Development, January 12, 2002; 129(23): 5409 - 5419. [Abstract] [Full Text] [PDF]

				N. Nesi, C. Jond, I. Debeaujon, M. Caboche, and L. Lepiniec The Arabidopsis TT2 Gene Encodes an R2R3 MYB Domain Protein That Acts as a Key Determinant for Proanthocyanidin Accumulation in Developing Seed PLANT CELL, September 1, 2001; 13(9): 2099 - 2114. [Abstract] [Full Text] [PDF]

				K. Sugimoto, S. Takeda, and H. Hirochika MYB-Related Transcription Factor NtMYB2 Induced by Wounding and Elicitors is a Regulator of the Tobacco Retrotransposon Tto1 and Defense-Related Genes PLANT CELL, December 1, 2000; 12(12): 2511 - 2528. [Abstract] [Full Text]

				E. Grotewold, M. B. Sainz, L. Tagliani, J. M. Hernandez, B. Bowen, and V. L. Chandler Identification of the residues in the Myb domain of maize C1 that specify the interaction with the bHLH cofactor R PNAS, November 22, 2000; (2000) 250379897. [Abstract] [Full Text]

				C. Spelt, F. Quattrocchio, J. N. M. Mol, and R. Koes anthocyanin1 of Petunia Encodes a Basic Helix-Loop-Helix Protein That Directly Activates Transcription of Structural Anthocyanin Genes PLANT CELL, September 1, 2000; 12(9): 1619 - 1632. [Abstract] [Full Text]

				K. Petroni, E. Cominelli, G. Consonni, G. Gusmaroli, G. Gavazzi, and C. Tonelli The Developmental Expression of the Maize Regulatory Gene Hopi Determines Germination-Dependent Anthocyanin Accumulation Genetics, May 1, 2000; 155(1): 323 - 336. [Abstract] [Full Text]

				J. B. Hollick, G. I. Patterson, I. M. Asmundsson, and V. L. Chandler Paramutation Alters Regulatory Control of the Maize pl Locus Genetics, April 1, 2000; 154(4): 1827 - 1838. [Abstract] [Full Text]

				W. Bruce, O. Folkerts, C. Garnaat, O. Crasta, B. Roth, and B. Bowen Expression Profiling of the Maize Flavonoid Pathway Genes Controlled by Estradiol-Inducible Transcription Factors CRC and P PLANT CELL, January 1, 2000; 12(1): 65 - 80. [Abstract] [Full Text] [PDF]

				F. Quattrocchio, J. Wing, K. van der Woude, E. Souer, N. de Vetten, J. Mol, and R. Koes Molecular Analysis of the anthocyanin2 Gene of Petunia and Its Role in the Evolution of Flower Color PLANT CELL, August 1, 1999; 11(8): 1433 - 1444. [Abstract] [Full Text]

				D. A. Selinger and V. L. Chandler A Mutation in the pale aleurone color1 Gene Identifies a Novel Regulator of the Maize Anthocyanin Pathway PLANT CELL, January 1, 1999; 11(1): 5 - 14. [Abstract] [Full Text]

				Y. Liu, L. Wang, J. L. Kermicle, and S. R. Wessler Molecular Consequences of Ds Insertion Into and Excision From the Helix-Loop-Helix Domain of the Maize R Gene Genetics, December 1, 1998; 150(4): 1639 - 1648. [Abstract] [Full Text]

				K. B. Singh Transcriptional Regulation in Plants: The Importance of Combinatorial Control Plant Physiology, December 1, 1998; 118(4): 1111 - 1120. [Full Text]

				M. Louis Lesnick and V. Lynn Chandler Activation of the Maize Anthocyanin Gene a2 Is Mediated by an Element Conserved in Many Anthocyanin Promoters Plant Physiology, June 1, 1998; 117(2): 437 - 445. [Abstract] [Full Text]

				C. B. Taylor Factories of the Future? Metabolic Engineering in Plant Cells PLANT CELL, May 1, 1998; 10(5): 641 - 644. [Full Text] [PDF]

				E. Grotewold, M. B. Sainz, L. Tagliani, J. M. Hernandez, B. Bowen, and V. L. Chandler Identification of the residues in the Myb domain of maize C1 that specify the interaction with the bHLH cofactor R PNAS, December 5, 2000; 97(25): 13579 - 13584. [Abstract] [Full Text] [PDF]