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Expression profiling of the Arabidopsis Mediator complex mutant pft1/med25 and wildtype infected with Fusarium oxysporum

ABSTRACT: The Mediator complex is an evolutionary conserved multiprotein complex that plays an essential role in initiating and regulating transcription. Its function is to act as a universal adaptor between RNA Polymerase II and DNA-bound transcription factors to translate regulatory information from activators and repressors to the transcriptional machinery. We have found that the PFT1 gene (which encodes the MED25 subunit of the Mediator complex) is required for the uncompromised expression of both salicylic acid- and jasmonate-dependent defense genes as well as resistance to the leaf-infecting fungal pathogens, Alternaria brassicicola and Botrytis cinerea in Arabidopsis. Surprisingly, we found that the pft1/med25 mutant showed increased resistance to the root infecting pathogen Fusarium oxysporum and that this resistance was independent of classical defense genes. In addition, the over-expression of PFT1 led to increased susceptibility to F. oxysporum. Therefore, to explore this phenomenon further, we wished to use whole genome transcript profiling to identify which genes may be playing a role in pft1/med25-mediated resistance to F. oxysporum.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE15236 | GEO | 2009/08/24

SECONDARY ACCESSION(S): PRJNA116431

REPOSITORIES: GEO

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Transcription profiling of Arabidopsis Mediator complex mutant pft1/med25 and wildtype infected with Fusarium oxysporum

Project description:The Mediator complex is an evolutionary conserved multiprotein complex that plays an essential role in initiating and regulating transcription. Its function is to act as a universal adaptor between RNA Polymerase II and DNA-bound transcription factors to translate regulatory information from activators and repressors to the transcriptional machinery. We have found that the PFT1 gene (which encodes the MED25 subunit of the Mediator complex) is required for the uncompromised expression of both salicylic acid- and jasmonate-dependent defense genes as well as resistance to the leaf-infecting fungal pathogens, Alternaria brassicicola and Botrytis cinerea in Arabidopsis. Surprisingly, we found that the pft1/med25 mutant showed increased resistance to the root infecting pathogen Fusarium oxysporum and that this resistance was independent of classical defense genes. In addition, the over-expression of PFT1 led to increased susceptibility to F. oxysporum. Therefore, to explore this phenomenon further, we wished to use whole genome transcript profiling to identify which genes may be playing a role in pft1/med25-mediated resistance to F. oxysporum. Experiment Overall Design: We grew both WT and pft1/med25 plants for four weeks in soil. After four weeks we treated the plants by root-dipping in either water or a F. oxysporum spore suspension before replanting them back into soil. There were four independent replicates of each treatment and each replicate contained 20 plants each. The treatments were WT (mock): pft1 (mock): WT (infected): pft1 (infected). Each replicate (16 in total) was harvested after 48 hours and the resulting RNA was used for hybridization to an Affymetrix ATH1 chip.

2009-09-05 | E-GEOD-15236 | biostudies-arrayexpress

shRNA-mediated knockdown of ETV4 and MED25 in the prostate cell line PC3 reveals set of genes potentially coregulated by MED25 and ETV4

Project description:Our in vitro binding studies support a model whereby MED25 exhibits multivalent interactions with a subset of related ETS factors, ETV1/4/5. We hypothesize that the interaction would allow for coregulation of genes by ETV1/4/5 and MED25, acting perhaps to link the ETVs to the Mediator complex. To explore this possibility, we compared the genome occupancy for FLAG-tagged MED25 and ETV4 in the prostate cancer cell line PC3, which overexpresses ETV4. We also tested for relevance of MED25 and ETV4 binding to for gene expression in PC3s. We found a high degree of overlap in the FLAG-MED25 and ETV4 ChIPs datasets consistent with our model, and also identified a subset of target genes co-dependent on Med25 and ETV4.

2019-06-28 | GSE133447 | GEO

Genome-wide occupancy of FLAG-MED25 and ETV4 in PC3 cells

2019-06-28 | GSE133445 | GEO

An Unexpected Encounter: Respiratory Syncytial Virus Non-Structural Protein 1 Interacts With Mediator Subunit MED25

Project description:Human respiratory syncytial virus (RSV) is the leading cause of severe acute lower respiratory tract infections in infants worldwide. Non-structural protein NS1 of RSV modulates the host innate immune response by acting as an antagonist of type I and type III interferon (IFN) production and signaling in multiple ways. It is likely that NS1 performs this function by interacting with different host proteins. In order to obtain a comprehensive overview of NS1 interaction partners, we performed three complementary protein-protein interaction screens i.e. BioID, MAPPIT and KISS. The BioID proximity screen was performed during an RSV infection in A549 cells. MED25, a subunit of the Mediator complex, was identified in all 3 performed screening methods as a potential NS1 interacting protein. We confirmed the interaction between MED25 and RSV NS1 by co-immunoprecipitation, not only upon overexpression of NS1, but also with endogenous NS1 during RSV infection. We also demonstrate that the replication of RSV is enhanced in MED25 knockout A549 cells, suggesting a potential antiviral role of MED25 during RSV infection. Mediator subunits function as transcriptional coactivators and are involved in transcriptional regulation of their target genes. Therefore, the interaction between RSV NS1 and cellular MED25 might be beneficial during an RSV infection as this can affect host transcription and the host immune response to infection.

2022-09-09 | PXD034627 | Pride

Expression profiling of wild-type Arabidopsis and an activation-tagged jaz7-1D line.

2016-02-08 | GSE61884 | GEO

RNA-seq analysis of ASCL1-induced neuronal differentiation in WT and MED25 knockout Glioblastoma Stem Cells

Project description:Glioblastoma stem cells were infected with lentivirus expressing a control (gRNA-AAVS1) or MED25 targeting gRNA to assess the role of MED25 in the stem cell compartment of Glioblastoma stem cells. Cells were the subjected to doxycycline-induced ASCL1 overexpression (+DOX) to induced neuronal differentiation.

2023-06-01 | GSE166979 | GEO

Arabidopsis thaliana

Project description:Expression profiling of the Arabidopsis Mediator complex mutant pft1/med25 and wildtype infected with Fusarium oxysporum

| PRJNA116431 | ENA

Hypoxic gene expression in med25-1

Project description:Hypoxic gene expression in med25-1

| PRJNA542085 | ENA

Expression profiling of wild-type Arabidopsis and an activation-tagged jaz7-1D line.

Project description:Jasmonate (JA) signaling plays a key role in mediating both resistance and susceptibility to the root-infecting fungal pathogen Fusarium oxysporum. Within this system, the roles of the JA-signaling repressor gene family of JASMONATE ZIM-domain (JAZ) genes had not been investigated. By screening JAZ T DNA insertion lines for altered resistance or susceptibility to F. oxysporum, we identified a JAZ7 mutant (jaz7-1D) highly susceptible to F. oxysporum infection. Further analyses revealed jaz7-1D exhibits constitutively active JAZ7 expression, enhanced expression of JA-defense marker genes, and increased sensitivity to JA-inhibition of root elongation. To further explore altered JA-signaling and JA-responses in this mutant, we use whole transcriptome profiling of jaz7-1D versus wild-type (Col-0) plants after mock/control and JA treatment. We grew both WT and jaz7-1D plants for four weeks in soil. After four weeks the plants were treated with 1 mL of 5% MeJA (dissolved in 100% ethanol) or 1 mL of Mock solution (100% ethanol) by applying the solutions to a cotton ball attached to a tray cover which was then used to enclose individual trays. Each tray and cover was then sealed by two layers of opaque plastic bags and secured with masking tape. There were four independent replicates of each treatment and each replicate contained leaf tissue from 20 plants. Each replicate (16 in total) was harvested after 6 hours and the resulting RNA was used for hybridization to an Affymetrix ATH1 chip.

2016-02-08 | E-GEOD-61884 | biostudies-arrayexpress

Hypoxic gene expression in med25-1

Project description:Plants as obligate aerobic organisms are severely affected by flooding-induced oxygen restriction. As a consequence, they suffer from energy deficits which are counterbalanced by activation of anaerobic metabolism genes through subgroup VII ETHYLENE-RESPONSE FACTOR (ERFVII) transcription factors. Despite the crucial role of ERFVIIs in regulating transcript responses to hypoxia in multiple plant species, it is not understood how these factors mechanistically initiate hypoxic gene expression. Here, we show that the Arabidopsis ERFVIIs RELATED TO APETALA 2.12 (RAP2.12) and RAP2.2 recruit the Mediator complex, present in all higher eukaryotes, in order to convey stress signals to RNA polymerase II. This is achieved by specific interaction with Mediator subunit 25 (AtMED25). Both RAP2.12 and RAP2.2 require AtMED25 for full activity and subsequent induction of target hypoxia core genes. Our findings support an interplay of multiple coactivators under hypoxia and underline the flexible nature of the Mediator complex composition, which both help to meet specific cellular demands under (hypoxic) stress conditions.

2019-12-31 | GSE130962 | GEO

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