13 research outputs found
Arabidopsis SWI/SNF chromatin remodeling complex binds both promoters and terminators to regulate gene expression
ATP-dependent chromatin remodeling complexes
are important regulators of gene expression in Eukaryotes.
In plants, SWI/SNF-type complexes have
been shown critical for transcriptional control of
key developmental processes, growth and stress responses.
To gain insight into mechanisms underlying
these roles, we performed whole genome mapping
of the SWI/SNF catalytic subunit BRM in Arabidopsis
thaliana, combined with transcript profiling
experiments. Our data showthatBRM occupies thousands
of sites in Arabidopsis genome, most of which
located within or close to genes. Among identified direct
BRM transcriptional targets almost equal numbers
were up- and downregulated upon BRM depletion,
suggesting that BRM can act as both activator
and repressor of gene expression. Interestingly,
in addition to genes showing canonical pattern of
BRM enrichment near transcription start site, many
other genes showed a transcription termination sitecentred
BRM occupancy profile. We found that BRMbound
3� gene regions have promoter-like features,
including presence of TATA boxes and high H3K4me3
levels, and possess high antisense transcriptional
activity which is subjected to both activation and
repression by SWI/SNF complex. Our data suggest
that binding to gene terminators and controlling transcription
of non-coding RNAs is another way through
which SWI/SNF complex regulates expression of its
targets
ANGUSTIFOLIA3 Binds to SWI/SNF Chromatin Remodeling Complexes to Regulate Transcription during Arabidopsis
Genetic analysis of functional redundancy of BRM ATPase and ATSWI3C subunits of Arabidopsis SWI/SNF chromatin remodelling complexes
Arabidopsis SWI/SNF chromatin remodeling complex binds both promoters and terminators to regulate gene expression
ANGUSTIFOLIA3 binds to SWI/SNF chromatin remodeling complexes to regulate transcription during Arabidopsis leaf development
The transcriptional coactivator ANGUSTIFOLIA3 (AN3) stimulates cell proliferation during Arabidopsis thaliana leaf development, but the molecular mechanism is largely unknown. Here, we show 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 FACTOR2, CONSTANS-LIKE5 (COL5), HECATE1 (HEC1), and ARABIDOPSIS RESPONSE REGULATOR4 (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. Moreover, SWI/SNF ASSOCIATED PROTEIN 73B (SWP73B) is recruited by AN3 to the promoters 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
GA responses of the <i>brm-1</i> mutant.
<p>(A, B), Elongation of <i>brm-1</i> hypocotyls and roots in response to 1 µM GA<sub>4</sub>. Plants were grown on ½ MS medium for 8 days under long-days conditions in the presence or absence of 1 µM GA<sub>4</sub>. GA application caused considerable elongation of the hypocotyls, but had little effect on <i>brm-1</i> root growth. Bar = 5 mm. (B), Hypocotyl length of plants grown as in A. Presented data are the means of 12 measurements ± s.d. (C), Flowering of <i>brm-1</i> plants in response to exogenous gibberellins. Plants were grown in soil under short-day conditions and treated with 10 µM GA<sub>3</sub>. At least 15 plants of each line/condition were scored. Data are the means ± s.d. Asterisks indicate significant differences from the wild type plants (p<0.01).</p
Concentration of gibberellins in wild type and mutant lines.
<p>The values are ng/g dry weight (s.e.). They are the means of three biological replicates except for the GA<sub>12</sub> and GA<sub>9</sub> measurements in <i>ga1-3</i>, for which 2 replicates were used. n.d. – not determined.</p
BRM acts through distinct mechanisms to regulate GA-mediated responses.
<p>(A), Germination of the <i>brm-1</i> mutant on 10 µM PAC is rescued by the <i>triple della</i> mutation. The progeny of <i>brm-1/BRM</i> plants were analyzed 10 days after sowing. (B), Phenotypes of 3-week-old plants grown on 2.5 µM PAC. The <i>brm-1/3xdella</i> line shows an intermediate growth phenotype. Bar = 5 mm. (C), RT-qPCR analysis of relative transcript levels of the <i>OFP16, EXP5, CYS2</i> and <i>LTP2</i> genes in 18-d-old wild type, <i>brm-1</i>, <i>ga1-3</i>, <i>ga1-3/brm-1</i>, <i>ga1-3/3xdella</i> and <i>ga1-3/brm-1/3xdella</i> lines. Transcript levels in the wild type were set to 1. Data are the means ± s.d. of 3 biological replicates. (D), Model of the role of BRM in regulating the expression of GA-responsive genes. BRM positively regulates the <i>GA3ox1</i> and <i>SCL3</i> genes involved in GA biosynthesis and signaling, and probably through this influences the expression of many GA-responsive genes in the opposite manner to DELLA repressors. In addition, BRM seems to act on a subset of GA-responsive genes independently of DELLA repressors. Also in this case, the effect exerted by BRM is typically in the opposite direction to that of DELLAs and is observed both for genes up- and down-regulated by the SWI/SNF complex (blue and red lines, respectively).</p
Gene Ontology (GO) categories statistically over-represented among genes differentially expressed in both <i>ga1-3</i> and <i>brm-1</i> mutants.
<p>Gene Ontology (GO) categories statistically over-represented among genes differentially expressed in both <i>ga1-3</i> and <i>brm-1</i> mutants.</p
<i>ga1-3/brm-1</i> mutant phenotypes.
<p>(A–B), Phenotypes of the <i>ga1-3</i>, <i>brm-1</i> and <i>ga1-3/brm-1</i> mutants grown on MS medium (18-d-old seedlings, A) or in soil (22-d-old plants, B). Bars = 10 mm. (C–F), Quantitative characterization of <i>brm-1</i>, <i>ga1-3</i> and <i>ga1-3/brm-1</i> mutants: root length of 18-d-old seedlings (C), rosette diameter at maturity (D) and flowering time under LD conditions (E). Data are the means ± s.d., 10 plants of each line were scored, except for <i>ga1-3/brm-1</i> (7 plants). * All <i>ga1-3/brm-1</i> plants except one failed to flower by the end of the experiment (80 days). (F), RT-qPCR analysis of relative transcript levels of <i>GA3ox1</i> and <i>SCL3</i> in 20-d-old wild type, <i>brm-1</i>, <i>ga1-3</i>, and <i>ga1-3/brm-1</i> lines. RT-qPCR data are the means ± s.d. of 3 biological replicates. Transcript levels in the wild type were set to 1.</p