Project description:ANGUSTIFOLIA 3 binds Arabidopsis SWI/SNF chromatin remodeling complexes to regulate transcription at the switch from leaf cell proliferation to expansion
Project description:The transcriptional coactivator ANGUSTIFOLIA 3 (AN3) stimulates cell proliferation during Arabidopsis leaf development, but the molecular mechanism is largely unknown. We show here that inducible nuclear localization of AN3 during initial leaf growth results in differential expression of important transcriptional regulators, including GROWTH REGULATING FACTORs (GRFs). Chromatin purification further revealed the presence of AN3 at the loci of GRF5, GRF6, CYTOKININ RESPONSE FACTOR 2 (CRF2), CONSTANS-LIKE 5 (COL5), HECATE 1 (HEC1), and ARABIDOPSIS RESPONSE REGULATOR 4 (ARR4). Tandem affinity purification of protein complexes using AN3 as bait identified plant SWITCH/SUCROSE NONFERMENTING (SWI/SNF) chromatin remodeling complexes formed around the ATPases BRAHMA (BRM) or SPLAYED (SYD). Moreover, SWI/SNF ASSOCIATED PROTEIN 73B (SWP73B) is recruited by AN3 to the promoter of GRF5, GRF3, COL5, and ARR4, and both SWP73B and BRM occupy the HEC1 promoter. Furthermore, we show that AN3 and BRM genetically interact. The data indicate that AN3 associates with chromatin remodelers to regulate transcription. In addition, modification of SWI3C expression levels increases leaf size, underlining the importance of chromatin dynamics for growth regulation. Our results place the SWI/SNF-AN3 module as a major player at the transition from cell proliferation to cell differentiation in a developing leaf.
Project description:The transcriptional coactivator ANGUSTIFOLIA 3 (AN3) stimulates cell proliferation during Arabidopsis leaf development, but the molecular mechanism is largely unknown. We show here that inducible nuclear localization of AN3 during initial leaf growth results in differential expression of important transcriptional regulators, including GROWTH REGULATING FACTORs (GRFs). Chromatin purification further revealed the presence of AN3 at the loci of GRF5, GRF6, CYTOKININ RESPONSE FACTOR 2 (CRF2), CONSTANS-LIKE 5 (COL5), HECATE 1 (HEC1), and ARABIDOPSIS RESPONSE REGULATOR 4 (ARR4). Tandem affinity purification of protein complexes using AN3 as bait identified plant SWITCH/SUCROSE NONFERMENTING (SWI/SNF) chromatin remodeling complexes formed around the ATPases BRAHMA (BRM) or SPLAYED (SYD). Moreover, SWI/SNF ASSOCIATED PROTEIN 73B (SWP73B) is recruited by AN3 to the promoter of GRF5, GRF3, COL5, and ARR4, and both SWP73B and BRM occupy the HEC1 promoter. Furthermore, we show that AN3 and BRM genetically interact. The data indicate that AN3 associates with chromatin remodelers to regulate transcription. In addition, modification of SWI3C expression levels increases leaf size, underlining the importance of chromatin dynamics for growth regulation. Our results place the SWI/SNF-AN3 module as a major player at the transition from cell proliferation to cell differentiation in a developing leaf. AN3-GR and wild-type (Col-0) plants were grown in vitro for 8 days and subsequently transferred to dexamethasone-containing medium for 8 hours. Developing leaves 1&2 of AN3-GR and wild-type plants were micro-dissected, and RNA was extracted. RNA from three biological repeats of dexamethasone-treated AN3-GR and dexamethasone-treated wild-type leaves was hybridized to Affymetrix ATH1 microarrays.
Project description:SWP73 subunits of SWI/SNF chromatin remodeling complexes (CRCs) are involved in key developmental pathways in Arabidopsis. We found, using microarray that inactivation of SWP73B caused altered expression of genes belonging to various regulatory pathways, including leaf and flower development. On the basis of this experiment and our other data we concluded that SWP73B modulates major developmental pathways.
Project description:Chromatin, in addition to its purely structural functions, is considered a major regulatory system coordinating various genetic networks in eukaryotes. Constant changes of gene expression programs are especially important for plants, which have to respond to environment by modulating their growth and development during whole lifetime. External and developmental signals can be transmitted through signaling cascades to chromatin remodeling complexes like SWI/SNF, which alter chromatin structure by moving, ejecting or restructuring nucleosomes. Genetic studies in Arabidopsis thaliana revealed that SWI/SNF chromatin remodeling complexes are critical for proper plant development and growth. Especially, BRM, a catalytic subunit of the complex, was shown to directly regulate several genes with important functions in leaf development, flowering initiation, as well as gibberellin and abscisic acid signaling. In this study, we profiled BRM global binding regions in Arabidopsis genome by ChIP-chip analysis. We found that BRM can bind to thousands of genes, many of which have key functions in hormone and stress signaling.
Project description:We identify three classes of Arabidopsis SWI/SNF chromatin remodeling complexes and demonstrate that they regulate different developmental processes by affecting chromatin accessibility.
Project description:We identify three classes of Arabidopsis SWI/SNF chromatin remodeling complexes and demonstrate that they regulate different developmental processes by affecting chromatin accessibility.
Project description:We identify three classes of Arabidopsis SWI/SNF chromatin remodeling complexes and demonstrate that they regulate different developmental processes by affecting chromatin accessibility.
Project description:Chromatin, in addition to its purely structural functions, is considered a major regulatory system coordinating various genetic networks in eukaryotes. External and developmental signals can be transmitted through signaling cascades to chromatin remodeling complexes like SWI/SNF, which alter chromatin structure by moving, ejecting or restructuring nucleosomes. Genetic studies in Arabidopsis thaliana revealed that SWI/SNF chromatin remodeling complexes are critical for proper plant development and growth. Especially, BRM, a catalytic subunit of the complex, was shown to directly regulate several genes with important functions in leaf development, flowering initiation, as well as gibberellin and abscisic acid signaling. In addition, SWI/SNF complexes were also shown to be involved in regulation of enhancer activity in animal cells and RNA-mediated transcriptional silencing in plants. In this study we performed transcript profiling of WT and brm-1 mutant using Agronomics microarrays for detecting changes in expression of both sense and antisense transcripts.
Project description:SWI/SNF chromatin remodeling complexes control gene expression by regulating chromatin structure. However, the full subunit composition of SWI/SNF complexes in plants remains unclear. Here we show that BRAHMA Interacting Protein 1 (BRIP1) and BRIP2 in Arabidopsis thaliana are core subunits of plant SWI/SNF complexes. BRIP1 and 2 are two homolog proteins. brip1 brip2 double mutants exhibit developmental phenotypes and a transcriptome strikingly similar to those of BRAHMA (BRM) mutants. Genetic interaction tests indicated that BRIP1 and 2 act together with BRM to regulate gene expression. Furthermore, BRIP1 and 2 physically interact with BRM-containing SWI/SNF complexes, and extensively co-localize with BRM at endogenous genes. Loss-of-brip1brip2 results in decreased BRM occupancy at almost all BRM target genes and substantially reduced subunits incorporation into the BRM-containing SWI/SNF complexes. Together, our work identifies new core subunits of BRM-containing SWI/SNF complexes in plants, and uncovers the essential role of these subunits in regulating the integrity (assembly) of SWI/SNF complexes in plants.