Arabidopsis thaliana
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA264533 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR1618855_1.fastq.gz | Fastqsanger.gz | |||
| SRR1618855_2.fastq.gz | Fastqsanger.gz | |||
| SRR1618856_1.fastq.gz | Fastqsanger.gz | |||
| SRR1618856_2.fastq.gz | Fastqsanger.gz | |||
| SRR1618857_1.fastq.gz | Fastqsanger.gz |
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Project description:Environment-dependent regulation of spliceosome activity by the LSM2-8 complex in Arabidopsis
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Project description:The assembly of the mitochondrial Complex I requires the expression of nuclear and mitochondrial located genes, co-factor biosynthesis and the assembly of at least 44 subunits. This requires the involvement of assembly factors that interact with subunits of Complex I but are not part of the final holocomplex. A novel plant specific mitochondrial matrix protein encoded by At1g76060, named COMPLEX I ASSEMBLY FACTOR 1 (CIAF1), containing a conserved LYR domain, was shown to be required for Complex I activity. T-DNA insertion mutants of At1g76060 lack the monomeric Complex I and the Supercomplex I+III, displaying the common Complex I growth deficient phenotype. Assembly of Complex I is stalled at 650 and 800 kDa intermediates in mitochondria isolated from these mutants. Evidence points to CIAF1 playing an essential role in the assembly of the peripheral matrix arm Complex I subunits to form the holoenzyme Complex I. We here performed quantitative proteomics by HiRIEF LC-MS with a TMT10plex isobaric tag labeling strategy comparing the control plant Col-0 and several mutant lines, including the mutant with inactivating insertion in At1g76060 (a.k.a. Ciaf1-1, a.k.a. SALK_143656). Among up-regulated proteins were components required for mitochondrial biogenesis, which points to a mitochondrial retrograde signaling pathway being activated and executed in response to the lack of Complex I.
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Project description:Pre-mRNA splicing is a highly regulated process catalyzing intron excision by spliceosome. Spliceosome activation is a major control step requiring dramatic protein and RNA rearrangements leading to a catalytically active complex. Prior research has linked hyperphosphorylation of SF3B1, a subunit of U2 snRNP, with spliceosome activation and catalytically active spliceosome, rendering a relevant kinase a key player for pre-mRNA splicing. Here we use OTS964, the first potent inhibitor of cyclin-dependent kinase 11 (CDK11), to show rapid and selective dephosphorylation of SF3B1 on threonines required for spliceosome activation. CDK11 associates with SF3B1 and its inhibition causes massive intron retention, block in precatalytic spliceosome complex B to activated spliceosome complex Bact transition and accumulation of non-functional spliceosomes on pre-mRNA and chromatin. These studies reveal crucial regulatory role of CDK11 in human pre-mRNA splicing and define the compound OTS964 as a quality chemical biology probe for CDK11.
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Project description:Recruitment of GSH into the nucleus during cell proliferation
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Project description:Polymerase-IV occupancy at RNA-directed DNA methylation sites requires SHH1
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Project description:Pre-mRNA splicing requires assembly of spliceosome that consists of hundreds of factors forming various dynamic complexes with or without small nuclear RNAs (snRNAs). Systematic identification of the splicing factors remains a significant challenge especially in vivo. In our genetic screening for the factors required for division asynchrony during Caenorhabditis elegans embryogenesis, we identified a highly conserved but uncharacterized essential protein, GAD-1, that is necessary for setting cell cycle length of intestine progenitors, a function that is shared with many other factors involved in transcription, pre-mRNA splicing or polyadenylation, suggesting its potential role in mRNA biogenesis. Co-immunoprecipitation followed by mass spectrometry reveals that GAD-1 mainly interacts with non-snRNP type of splicing complex called NineTeen Complex (NTC). Consistent with this, RNA-seq analysis demonstrates pervasive defects in pre-mRNA splicing in gad-1 mutants. Transgenic reporter assay shows its ubiquitous and nuclear expression across developmental stages. Immunostaining of the C-terminal domain of RNA polymerase II demonstrates a GAD-1’s role in transcription elongation. In agreement with this, depletion of GAD-1 and its interacting partners inhibits expression of both ubiquitous and tissue-specific genes, supporting that both GAD-1 and many of its interacting proteins are novel components of NTC or its associated spliceosome. Taken together, we identify GAD-1 and its multiple interacting partners as novel components of spliceosome in vivo through which they regulate pre-mRNA splicing and transcription elongation.
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Project description:SKIP Is a Component of the Spliceosome Linking Alternative Splicing and the Circadian Clock in Arabidopsis
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Project description:Shoot apical meristem (SAM) of higher plant composed of a few distinct cell types. All the cells in a mature plant’s SAM derived from 30~35 stem cells reservoir which are located at the tip of the apex. Plants ability to give rise diverse cell types from a pool of pluripotent stem cells requires orchestrated gene network that controls the cell fate commitment during the meristem development. To understand, how gene regulatory networks control cell identities switches during cell differentiation requires resolution in recording their gene expression pattern at single cell resolution. An earlier expression map involving three-cell population of stem cell niche revealed complex expression pattern among the cell types1. We developed this approach further and report here a gene expression map using cell-sorting methods for fluorescent protein marked cells in Arabidopsis shoot. The map covered 10 cell populations. This gene expression map represents data from 10 different cell types from Arabidopsis SAM. It will be first step in defining the function of many unknown genes in model plant Arabidopsis. Based on the in situ hybridization we identified 7 new cell types specific gene expression patterns. The promoters of these genes were used to generate fluorescent reporters. After treating the SAM with protoplasting cocktail, we sorted the fluorescent protein tagged cells using fluorescent activated cell sorter (FACS). The purified cell population was used to isolate RNA. Two round of RNA amplification was performed before microarray hybridization.
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