FIONA1-mediated methylation of the 3'UTR of FLC affects FLC transcript levels and flowering in Arabidopsis
Ontology highlight
ABSTRACT:
ORGANISM(S): Arabidopsis thaliana
PROVIDER: GSE212766 | GEO | 2022/09/12
REPOSITORIES: GEO
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Project description:Adenosine bases of RNA can be transiently modified by the deposition of a methyl-group to form N6-methyladenosine (m6A). This adenosine-methylation is an ancient process and the enzymes involved are evolutionary highly conserved. A genetic screen designed to identify suppressors of late flowering transgenic Arabidopsis plants overexpressing the miP1a microProtein yielded a new allele of the FIONA1 (FIO1) m6A-methyltransferase. To characterize the early flowering phenotype of fio1 mutant plants we employed an integrative approach of mRNA-seq, Nanopore direct RNA-sequencing and meRIP-seq to identify differentially expressed transcripts as well as differentially methylated RNAs. We provide evidence that FIO1 is the elusive methyltransferase responsible for the 3'-end methylation of the FLOWERING LOCUS C (FLC) transcript. Furthermore, our genetic and biochemical data suggest that 3'-methylation stabilizes FLC mRNAs and non-methylated FLC is a target for rapid degradation.
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Project description:FIONA1-mediated methylation of the 3'UTR of FLC affects FLC transcript levels and flowering in Arabidopsis
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Project description:Plants of three different genotypes (FRI FLC, FRI flc and fri flc) were induced to flowering by shifting from short day conditions to long day conditions. FRI=FRIGIDA, FLC=FLOWERING LOCUS C.
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Project description:Several pathways control time to flowering in Arabidopsis thaliana through transcriptional and posttranscriptional gene regulation. In recent years, mRNA processing has gained interest as a critical regulator of flowering time control in plants. However, the molecular mechanisms linking RNA splicing to flowering time are not well understood. In a screen for Arabidopsis early flowering mutants we identified an allele of BRR2a. BRR2 proteins are components of the spliceosome and highly conserved in eukaryotes. Arabidopsis BRR2a is ubiquitously expressed in all analyzed tissues and involved in the processing of flowering time gene transcripts, most notably FLC. A missense mutation of threonine 895 in BRR2a caused defects in FLC splicing and greatly reduced FLC transcript levels. Reduced FLC expression increased transcription of FT and SOC1 leading to early flowering in both short and long days. Genome-wide experiments established that only a small set of introns was not correctly spliced in the brr2a mutant. Compared to control introns, retained introns were often shorter and GC-poor, had low H3K4me1 and CG methylation levels, and were often derived from genes with a high-H3K27me3-low-H3K36me3 signature. We propose that BRR2a is specifically needed for efficient splicing of a subset of introns characterized by a combination of factors including intron size, sequence and chromatin, and that FLC is most sensitive to splicing defects.
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Project description:Arabiposis plants with conbinations of different FRIGIDA (FRI) and FLOWERING LOCUS C (FLC) alleles grown in short days (9L:15D) for 30 days at 21°C, then shifted to long days (16L:8D). Genotypes: Columbia wild type (Col-0): fri FLC Columbia with introgressed FRI from Sf-2: FRI FLC Columbia with introgressed FRI and deleted FLC (flc-3): FRI flc Columbia with deleted FLC (flc-3): fri flc Time points: 0, 2, and 4 days after shift to long days Keywords = flowering Keywords: time-course
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Project description:Flowering time is a key adaptive and agronomic trait. In Arabidopsis, natural variation in expression levels of the floral repressor FLOWERING LOCUS C (FLC) leads to differences in vernalization. In Brassica napus there are nine copies of FLC. Here, we study how these multiple FLC paralogues determine vernalization requirement as a system. We collected transcriptome time series for Brassica napus spring, winter, semi-winter, and Siberian kale crop types. Modelling was used to link FLC expression dynamics to floral response following vernalization. We show that relaxed selection pressure has allowed expression of FLC paralogues to diverge, resulting in variation of FLC expression during cold treatment between paralogues and accessions. We find that total FLC expression dynamics best explains differences in cold requirement between cultivars, rather than expression of specific FLC paralogues. The combination of multiple FLC paralogues with different expression dynamics leads to rich behaviour in response to cold and a wide range of vernalization requirements in B. napus. We find evidence for different strategies to determine the response to cold in existing winter rapeseed accessions.
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Project description:The autonomous flowering pathway promotes flowering in rapid-flowering accessions of Arabidopsis thaliana by reducing the expression of the key flowering repressor gene FLC. Although previous studies identified several autonomous pathway components, little is known about how these components cooperate in the regulation of flowering time. Here, we identified an autonomous pathway complex (APC) composed of three known autonomous pathway components (FLD, LD, and SDG26) and three previously uncharacterized components (EFL2, EFL4, and APRF1). Loss-of-function mutations of all of these components result in increased FLC expression and delayed flowering. The late-flowering phenotype is independent of photoperiod and can be overcome by vernalization, supporting the inference that the complex regulates flowering time specifically through the autonomous pathway. Our ChIP-seq analyses indicated that the APC components are required for the suppression of the histone modifications (H3Ac, H3K4me3, and H3K36me3) associated with the activation of gene expression and for the promotion of the histone modification (H3K27me3) associated with the repression of gene expression at FLC. We also found that the C-terminal coiled-coil domain of SDG26 binds to DNA in vitro and that the DNA-binding ability mediates the association of SDG26 with FLC chromatin in vivo. These results reveal the molecular basis of the autonomous pathway.
2022-06-25 | GSE193252 | GEO
Project description:The autonomous flowering pathway promotes flowering in rapid-flowering accessions of Arabidopsis thaliana by reducing the expression of the key flowering repressor gene FLC. Although previous studies identified several autonomous pathway components, little is known about how these components cooperate in the regulation of flowering time. Here, we identified an autonomous pathway complex (APC) composed of three known autonomous pathway components (FLD, LD, and SDG26) and three previously uncharacterized components (EFL2, EFL4, and APRF1). Loss-of-function mutations of all of these components result in increased FLC expression and delayed flowering. The late-flowering phenotype is independent of photoperiod and can be overcome by vernalization, supporting the inference that the complex regulates flowering time specifically through the autonomous pathway. Our ChIP-seq analyses indicated that the APC components are required for the suppression of the histone modifications (H3Ac, H3K4me3, and H3K36me3) associated with the activation of gene expression and for the promotion of the histone modification (H3K27me3) associated with the repression of gene expression at FLC. We also found that the C-terminal coiled-coil domain of SDG26 binds to DNA in vitro and that the DNA-binding ability mediates the association of SDG26 with FLC chromatin in vivo. These results reveal the molecular basis of the autonomous pathway.
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Project description:Hight throughput techniques have revealed huge complexity in antisense RNAs in many organisms. We have explored the complexity of this class of transcripts and functional links to Polycomb silencing thought analysis of non-coding RNAs of Arabidopsis FLC. FLC is repressed and epigenetically silenced by prolonged cold, enabling plants to undergo the floral transition. Single nucleotide resolution tiling array revealed long non-coding transcripts covering the entire FLC locus. The most abundant of these are capped and polyadenylated, initiate over a 100 nucleotide window just downstream of the sense polyA site, are differentially spliced and terminate either within the sense gene or its promoter. Their levels correlate with FLC sense transcripts in all mutants and conditions tested except cold treatment. The antisense transcripts were strongly but transiently cold-induced, much earlier than other vernalization markers, and this coincided with reduction in sense FLC transcription but not sense FLC mRNA levels. Addition of the FLC antisense 5'/sense 3' region to a GFP transgene was sufficient to confer cold-induced silencing of thet fusion; however this silencing was not epigenetically manteined. These processes were all independent of the function of the Polycomb proteins required for maintenance of FLC silencing. Our data suggest that FLC antisense transcripts induce transient FLC transcriptional silencing, possibly through promoter interference, with the epigenetic silencing requiring subsequent recruitment of Polycomb machinery.
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