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1.
Plant Cell ; 27(6): 1605-19, 2015 Jun.
Article in English | MEDLINE | ID: mdl-26036253

ABSTRACT

Most molecular processes during plant development occur with a particular spatio-temporal specificity. Thus far, it has remained technically challenging to capture dynamic protein-protein interactions within a growing organ, where the interplay between cell division and cell expansion is instrumental. Here, we combined high-resolution sampling of the growing maize (Zea mays) leaf with tandem affinity purification followed by mass spectrometry. Our results indicate that the growth-regulating SWI/SNF chromatin remodeling complex associated with ANGUSTIFOLIA3 (AN3) was conserved within growing organs and between dicots and monocots. Moreover, we were able to demonstrate the dynamics of the AN3-interacting proteins within the growing leaf, since copurified GROWTH-REGULATING FACTORs (GRFs) varied throughout the growing leaf. Indeed, GRF1, GRF6, GRF7, GRF12, GRF15, and GRF17 were significantly enriched in the division zone of the growing leaf, while GRF4 and GRF10 levels were comparable between division zone and expansion zone in the growing leaf. These dynamics were also reflected at the mRNA and protein levels, indicating tight developmental regulation of the AN3-associated chromatin remodeling complex. In addition, the phenotypes of maize plants overexpressing miRNA396a-resistant GRF1 support a model proposing that distinct associations of the chromatin remodeling complex with specific GRFs tightly regulate the transition between cell division and cell expansion. Together, our data demonstrate that advancing from static to dynamic protein-protein interaction analysis in a growing organ adds insights in how developmental switches are regulated.


Subject(s)
Plant Leaves/growth & development , Plant Proteins/physiology , Zea mays/genetics , Conserved Sequence/genetics , Conserved Sequence/physiology , Plant Growth Regulators/genetics , Plant Growth Regulators/physiology , Plant Leaves/genetics , Plant Proteins/genetics , Tandem Mass Spectrometry
2.
Plant Physiol ; 164(3): 1350-64, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24488968

ABSTRACT

DNA methylation is an important and widespread epigenetic modification in plant genomes, mediated by DNA methyltransferases (DMTs). DNA methylation is known to play a role in genome protection, regulation of gene expression, and splicing and was previously associated with major developmental reprogramming in plants, such as vernalization and transition to flowering. Here, we show that DNA methylation also controls the growth processes of cell division and cell expansion within a growing organ. The maize (Zea mays) leaf offers a great tool to study growth processes, as the cells progressively move through the spatial gradient encompassing the division zone, transition zone, elongation zone, and mature zone. Opposite to de novo DMTs, the maintenance DMTs were transcriptionally regulated throughout the growth zone of the maize leaf, concomitant with differential CCGG methylation levels in the four zones. Surprisingly, the majority of differentially methylated sequences mapped on or close to gene bodies and not to repeat-rich loci. Moreover, especially the 5' and 3' regions of genes, which show overall low methylation levels, underwent differential methylation in a developmental context. Genes involved in processes such as chromatin remodeling, cell cycle progression, and growth regulation, were differentially methylated. The presence of differential methylation located upstream of the gene anticorrelated with transcript expression, while gene body differential methylation was unrelated to the expression level. These data indicate that DNA methylation is correlated with the decision to exit mitotic cell division and to enter cell expansion, which adds a new epigenetic level to the regulation of growth processes.


Subject(s)
DNA Methylation/genetics , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , Genes, Plant/genetics , Plant Leaves/growth & development , Plant Leaves/genetics , Zea mays/genetics , Base Sequence , Cell Division , Centromere/metabolism , Genetic Loci , Methyltransferases/metabolism , Phylogeny , Plant Leaves/enzymology , Polymerase Chain Reaction , Polymorphism, Genetic , Transcription, Genetic , Zea mays/cytology , Zea mays/enzymology , Zea mays/growth & development
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