The basic helix loop helix domain of maize R links transcriptional regulation and histone modifications by recruitment of an EMSY-related factor.
The control of anthocyanin accumulation in maize by the cooperation of the basic helix-loop-helix (bHLH) protein R with the MYB transcription factor C1 provides one of the best examples of plant combinatorial transcriptional control. Establishing the function of the bHLH domain of R has remained elusive, and so far no proteins that interact with this conserved domain have been identified. We show here that the bHLH domain of R is dispensable for the activation of transiently expressed genes yet is essential for the activation of the endogenous genes in their normal chromatin environment. The activation of A1, one of the anthocyanin biosynthetic genes, is associated with increased acetylation of histone 3 (H3) at K9/K14 in the promoter region to which the C1/R complex binds. We identified R-interacting factor 1 (RIF1) as a nuclear, AGENET domain-containing EMSY-like protein that specifically interacts with the bHLH region of R. Knockdown experiments show that RIF1 is necessary for the activation of the endogenous promoters but not of transiently expressed genes. ChIP experiments established that RIF1 is tethered to the regulatory region of the A1 promoter by the C1/R complex. Together, these findings describe a function for the bHLH domain of R in linking transcriptional regulation with chromatin functions by the recruitment of an EMSY-related factor.
Hernandez
J Marcela
JM
Ohio State Biochemistry Program, Ohio State University, Columbus, OH 43210, USA.
Feller
Antje
A
Morohashi
Kengo
K
Frame
Kenneth
K
Grotewold
Erich
E
D000872:Anthocyanins / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D006657:Histones / Q000378:metabolism*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D014158:Transcription, Genetic; D003313:Zea mays / Q000235:genetics*
D000872:Anthocyanins; D051792:Basic Helix-Loop-Helix Proteins; D006657:Histones; D009687:Nuclear Proteins; D010940:Plant Proteins; C085200:R protein, Zea mays