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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gametophytic transcription factors

ABSTRACT: Following our initial transcriptomic analyses of the male gametophyte development (Honys and Twell, Genome Biol 5:R85, 2004), we identified several candidate genes for the function of transcriptional regulators of the male gametophyte development. Two transcription factors with verified function (bZIP and C2H2 ZF) were selected for the identification of their regulons. Mature pollen was isolated as published previously (Honys and Twell Plant Physiol 132:640-652, 2003) from Col-0 wild type plants and T-DNA insertion mutants (SAIL) homozygous for tested genes. 6 samples were used in this experiment.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Nottingham Arabidopsis Stock Centre (NASC)

PROVIDER: E-GEOD-20224 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Gametophytic transcription factors

Project description:Following our initial transcriptomic analyses of the male gametophyte development (Honys and Twell, Genome Biol 5:R85, 2004), we identified several candidate genes for the function of transcriptional regulators of the male gametophyte development. Two transcription factors with verified function (bZIP and C2H2 ZF) were selected for the identification of their regulons. Mature pollen was isolated as published previously (Honys and Twell Plant Physiol 132:640-652, 2003) from Col-0 wild type plants and T-DNA insertion mutants (SAIL) homozygous for tested genes.

2010-04-20 | GSE20224 | GEO

Gene expression in mature pollen and sperm cells versus young seedling as a vegetative sporophyte reference control

Project description:Sperm cells represent the male partner that fuses with the egg cell during fertilization in all multi-cellular eukaryotic organisms, and, in flowering plants, is a founder of both embryo and nutritive endosperm. We examined the transcriptome of Oryza sativa ssp. japonica using the Affymetrix 57K rice genome GeneChip to provide an overview of genes activated in the paternal gamete. We used microarrays to detail the global program of gene expression in mature pollen and sperm cells at anthesis, which constitutes the stage immediately preceding pollen germination and gamete deposition leading up to fertilization. This documents the condition of the male gametophytic cells prior to fertilization. Keywords: tissue expression profile, male gametophyte, sperm, pollen, seedling Male gametophytic cells consisting of isolated pollen and isolated sperm cells collected at anthesis were compared with seedlings containing leaf, stem, root and apical meristems as a vegetative sporophytic control; three separate biological replicates were used.

2012-04-29 | E-GEOD-17002 | biostudies-arrayexpress

Characterization of the small RNA populations of developing Arabidopsis thaliana pollen.

Project description:Small RNA diversity and function has been widely characterized in various tissues of the sporophytic generation of the angiosperm model Arabidopsis thaliana. In contrast, there is limited knowledge about small RNA diversity and their roles in developing male gametophytes. We thus carried out small RNA sequencing on RNA isolated from four stages of developing Arabidopsis thaliana pollen. Spores from 4 stages of pollen development (UNM: Uninucleate microspore M-bM-^@M-^S BCP: Bicellular pollen M-bM-^@M-^S TCP: Tricellular pollen M-bM-^@M-^S MP: Mature pollen) were isolated using a percoll gradient-based method (Honys and Twell, 2004) and the small RNA fraction for each sample was isolated and sequenced by Illumina technology. Reference: Honys, D. and Twell, D. (2004) Transcriptome analysis of haploid male gametophyte development in Arabidopsis. Genome Biol. 5/11/R85.

2014-07-01 | E-GEOD-34190 | biostudies-arrayexpress

Expression data from fission yeast deletion mutant of Klf1, a C2H2 zinc finger-transcription factor, in G0 and VE phases

Project description:Fission yeast Schizosaccharomyces pombe is a model for studying cellular quiescence. Shifting to medium without a nitrogen-source induces proliferative cells to enter long-term G0 quiescence. Klf1 is a Kruppel-like transcription factor with a 7-amino acid-spaced C2H2-type zinc finger motif. The deletion mutant M-bM-^HM-^Fklf1 normally divides in vegetative medium, but proliferation is not restored after long-term G0 quiescence. Cell biologic, transcriptomic, and metabolomic analyses revealed a unique phenotype of the M-bM-^HM-^Fklf1 mutant in quiescence. Mutant cells had diminished transcripts related to signaling molecules for switching to differentiation. In contrast, proliferative metabolites for cell-wall assembly and antioxidants significantly increased. Further, the size of the M-bM-^HM-^Fklf1 cells is markedly increased during quiescence due to the aberrant accumulation of calcofluor-positive chitin-like materials beneath the cell wall. After 4 weeks quiescence, the ability for reversible proliferation is lost, but energy metabolism is maintained. Klf1 thus plays a role in G0 phase longevity through enhancing the differentiation signal and suppressing metabolism for growth. If Klf1 is lost, S. pombe fails to maintain a constant cell size during quiescence. Gene expression profile of fission yeast wild type cells and klf1-gene disruptant cells in proliferating state and in quiescent state. Type of experiment: Comparing between wild type cells and klf1-gene disruptant cells in proliferating condition and in quiescent condition. Experimental factor: Exponentially proliferating state in synthetic medium, EMM2, and quiescent G0 state in nitrogen-depleted synthetic medium, EMM2-N. Quality control steps taken: All experiments were repeated twice in each condition.

2013-12-01 | E-GEOD-50986 | biostudies-arrayexpress

Large-scale characterization of DNA binding landscape of eukaryotic C2H2 zinc fingers

Project description:This SuperSeries is composed of the SubSeries listed below. Cys2-His2 zinc finger (C2H2-ZF) proteins represent the largest class of putative human transcription factors (TFs). However, it is unknown whether most C2H2-ZFs even bind DNA, or what sequences they bind. Using a combination of bacterial one-hybrid (B1H) assays, protein-binding microarrays (PBMs), and ChIP-seq, we have found that most natural C2H2-ZFs bind DNA both in vitro and in vivo. This SuperSeries contains the data for identification of C2H2-ZF binding preferences using these three approaches. Refer to individual Series

2015-02-17 | E-GEOD-52523 | biostudies-arrayexpress

The C2H2-ZF transcription factor Zfp335 recognizes two consensus motifs using separate zinc finger arrays

Project description:The complexities of DNA recognition by transcription factors with multiple Cys2-His2 zinc fingers (C2H2-ZFs) remain poorly studied. We previously reported a mutation (R1092W) in the C2H2-ZF transcription factor Zfp335 which led to selective loss of binding at a subset of targets, although the basis for this was unclear. We show that Zfp335 binds DNA and drives transcription via recognition of two distinct consensus motifs by separate ZF clusters, and identify the specific motif interaction disrupted by R1092W. Our work presents Zfp335 as a model for understanding how C2H2-ZF TFs may utilize multiple recognition motifs to control gene expression.

2016-07-11 | GSE83116 | GEO

Large-scale characterization of DNA binding landscape of eukaryotic C2H2 zinc fingers

2015-02-17 | GSE52523 | GEO

Expression data from wild-type and rph1-deleted yeasts

Project description:Cells respond to environmental signals by alteration of gene expression through action of transcription factors. A JmjC-domain-containing protein Rph1 belongs to the C2H2 zinc finger protein family and functions to repress transcription of PHR1 via histone demethylation. However, additional targets of Rph1 remain largely unknown. Here, we investigate the regulatory network of Rph1 by microarray analyses. More than 75% of Rph1-regulated genes showed increased expression in rph1M-NM-^T, suggesting Rph1 serves as a transcriptional repressor under physiological conditions. The binding motif of Rph1 resembling the STress Response Element (STRE) was over-represented in the promoters of Rph1-repressed genes. In addition, significant proportions of Rph1-regulated genes responded to DNA damage and environmental stress, implying a repressive role of Rph1 in the cross-protection of stress responses. We used expression microarray to identify Rph1-regulated genes and established Rph1 functioned as a transcriptional repressor to link DNA damage signaling and general stress response. We compared gene expression in wild-type and rph1-deleted strains cultured in rich medium (YPD) during exponential growth. 3 biological samples were analyzed.

2013-11-06 | E-GEOD-39950 | biostudies-arrayexpress

Highly parallel identification of sequence preferences for eukaryotic C2H2 zinc finger domains using a bacterial 1-hybrid assay

Project description:The largest and most diverse class of eukaryotic transcription factors contain Cys2-His2 zinc fingers (C2H2-ZFs), each of which typically binds a DNA nucleotide triplet within a larger binding site. Frequent recombination and diversification of their DNA-contacting residues suggests that these zinc fingers play a prevalent role in adaptive evolution. Very little is known about the function and evolution of the vast majority of C2H2-ZFs, including whether they even bind DNA. Using the bacterial 1-hybrid (B1H) system, we determined DNA-binding motifs for thousands of individual natural C2H2-ZFs, and correlated them with the C2H2-ZF specificity residues. We generated 47,072 variants of the DNA-binding domain of the well-characterized three-C2H2-ZF protein Egr1, in which the third C2H2-ZF ('F3') was replaced with a natural C2H2-ZF. We screened this library using the bacterial 1-hybrid (B1H) system to assess the relative preference of each protein for 200 of the 256 possible NNN NGG GCG variants of the optimal Egr1 binding site, GCG TGG GCG, each in duplicate. Each sample might have been sequenced multiple times, in which case multiple raw and processed data files are indicated. This data represent bacterial 1-hybrid assays using three different "Original Library Samples", OLS-A, OLS-B, and OLS-C. The 'characteristics:OLS' indicates which OLS was used as the initial material for each of the assays. The processed data contains unique ID given to each sequence in the library and its abundance count. The 'OLS.info.xlsx' file contains the mapping of these IDs to different sequences.

2015-02-17 | E-GEOD-52521 | biostudies-arrayexpress

Evaluation of DNA-binding preferences of non-metazoan C2H2 zinc finger proteins by PBM

Project description:The largest and most diverse class of eukaryotic transcription factors contain Cys2-His2 zinc fingers (C2H2-ZFs). Here, we have explored the diversity of sequence preferences of C2H2-ZF proteins in non-metazoan organisms using PBM experiments.

2017-07-10 | GSE73117 | GEO

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