Protein idetification by using DNA pull-down & MS assay
Ontology highlight
ABSTRACT:
INSTRUMENT(S):
ORGANISM(S): Pseudomonas Syringae Pv. Actinidiae Str. Shaanxi_m228
TISSUE(S): Prokaryotic Cell
DISEASE(S): Disease Free
SUBMITTER: 
Zhibo Zhao
LAB HEAD: Zhibo Zhao
PROVIDER: PXD023722 | Pride | 2021-03-23
REPOSITORIES: Pride
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| 20190821-002-DUI-5.wiff | Wiff | |||
| 20190821-002-DUI-5.wiff.scan | Wiff | |||
| 20190821-002-SHI-4.wiff | Wiff | |||
| 20190821-002-SHI-4.wiff.scan | Wiff | |||
| M228gp_aa_database.fas | Other |
Items per page: 5 1 - 5 of 6 |
Similar Datasets
Project description:Eukaryotic topoisomerase 1 (TOP1) regulates DNA topology to ensure efficient DNA replication and transcription. TOP1 is also a major driver of endogenous genome instability, particularly when its catalytic intermediate - a covalent TOP1-DNA adduct known as a TOP1 cleavage complex (TOP1cc) - is stabilised. TOP1ccs are highly cytotoxic and a failure to resolve them underlies the pathology of neurological disorders but is also exploited in cancer therapy where TOP1ccs are the target of widely used frontline anti-cancer drugs. A critical enzyme for TOP1cc resolution is the tyrosyl-DNA phosphodiesterase, TDP1, which hydrolyses the bond that links a tyrosine in the active site of TOP1 to a 3’ phosphate group on a single-stranded (ss)DNA break. However, TDP1 can only process small peptide fragments from ssDNA ends, raising the question of how the ~90 kDa TOP1 protein is processed upstream of TDP1. We find that TEX264 fulfils this role by forming a complex with the p97 ATPase and the SPRTN metalloprotease. We show that TEX264 recognises both unmodified and SUMO1-modifed TOP1 and initiates TOP1cc repair by recruiting p97 and SPRTN. TEX264 localises to the nuclear periphery, associates with DNA replication forks, and counteracts TOP1ccs during DNA replication. Altogether, our study elucidates the existence of a specialised repair complex required for upstream proteolysis of TOP1ccs and their subsequent resolution.
2020-03-13 | PXD017239 | Pride
Project description:affy_syringae_malaga_athaliana - affy_syringae_malaga_athaliana - - Pseudomonas syringae is a plant pathogenic bacterium that relies on a type III secretion system (T3SS) to cause disease in susceptible hosts and to trigger defence in resistant plants. The T3SS translocates effector proteins directly inside the plant cell. Some P. syringae pathovars translocate the effector HopZ1a, which is recognized in Arabidopsis, triggering the hypersensitive response (HR). This resistance does not depend on the usual defence pathways involved in resistance against other avirulence effectors, so it would be very helpful to know the transcriptomic events that take place in the context of a hypersensitive response triggered by HopZ1a. ULP genes codifies for plant SUMO-proteases. Recent studies by our group revealed that they could be involved in plant defense against microorganisms.-- We infiltrated Arabidopsis thaliana ecotype Columbia wild type with 10mM MgCl2 (as mock inoculation), the virulent pathogen Pseudomonas syringae pv. tomato (Pto), Pto expressing the effector HopZ1a and Pto expressing a HopZ1a derivative carrying a point mutation in a residue essential for the cystein-protease activity. We also grew ulp1c/ulp1d mutant plants to analyze them either without treatment or inoculated with MgCl2 (as mock) and Pto wt. Keywords: gene knock out,normal vs antisens mutant comparison 24 arrays - ATH1
2011-04-20 | E-GEOD-21920 | biostudies-arrayexpress
Project description:Our previous study on CBP60g, a calmodulin binding protein that is important for disease resistance and microbe-associated molecular pattern (MAMP)-induced SA accumulation, led to our discovery of a closely related family member CBP60h. CBP60h is also important for defense against P. syringae but is induced differently by pathogen and MAMP stimulus. Transcriptome profiling of cbp60h mutants suggested that CBP60h might be primarily functioning in response against P. syringae. We constructed a double mutant of cbp60g and cbp60h, which demonstrated severely defective defense against P. syringae and SA accumulation. Profiling of the cbp60g/h showed that its expression pattern is very similar to that of pad4. Transient expression in Tobacco showed that both CBP60g and CBP60h localized to nucleus. Our observation suggest that CBP60g and CBP60h share partially redundant but critical role in defense response and SA signaling. This experiment consists of three biological replicates. For each genotype, two leaves per plant were pooled from three pots to prepare total RNA.
2009-11-03 | E-GEOD-18865 | biostudies-arrayexpress
Project description:Salicylic acid (SA)-induced defense responses are important factors during effector triggered immunity and microbe-associated molecular pattern (MAMP)-induced immunity in plants. This article presents evidence that a member of the Arabidopsis CBP60 gene family, CBP60g, contributes to MAMP-triggered SA accumulation. CBP60g is inducible by both pathogen and MAMP treatments. Pseudomonas syringae growth is enhanced in cbp60g mutants. Expression profiles of a cbp60g mutant after MAMP treatment are similar to those of sid2 and pad4, suggesting a defect in SA signaling. Accordingly, cbp60g mutants accumulate less SA when treated with the MAMP flg22 or a P. syringae hrcC strain that activates MAMP signaling. MAMP-induced production of reactive oxygen species and callose deposition are unaffected in cbp60g mutants. CBP60g is a calmodulin-binding protein with a calmodulin-binding domain located near the N-terminus. Calmodulin binding is dependent on Ca2+. Mutations in CBP60g that abolish calmodulin binding prevent complementation of the SA production and bacterial growth defects of cbp60g mutants, indicating that calmodulin binding is essential for the function of CBP60g in defense signaling. These studies show that CBP60g constitutes a calmodulin-dependent link between MAMP recognition and SA accumulation that is important for resistance to P. syringae. This experiment consists of three biological replicates. For each genotype, two leaves per plant were pooled from three pots to prepare total RNA.
2010-05-07 | E-GEOD-14237 | biostudies-arrayexpress
Project description:To further investigate the transcription cofactor of Smad3 that regulated the transcription of PRDM16, we conducted DNA pull-down assay. we treated HK-2 cells with TGF-β and extracted the nuclear proteins. And the nuclear proteins were incubated with streptavidin magnetic beads, and biotin-labeled DNA probes which bound to the promoter of PRDM16 specifically. After repeatedly washing, the pull-down proteins were analyzed by LC-MS/MS proteomics.
2025-07-14 | PXD044753 | Pride
Project description:Expression profiling of wild-type plants and mutants with defects in key components of the defense signaling network was used to model the Arabidopsis network 24 hours after infection by Pseudomonas syringae pv. maculicola strain Psm ES4326. Results using the Affymetrix ATH1 array revealed that expression levels of most pathogen-responsive genes were affected by mutations in coi1, ein2, npr1, pad4, or sid2. These five mutations defined a small number of different expression patterns displayed by the majority of pathogen-responsive genes. P. syringae pv. tomato strain Pst DC3000 elicited a much weaker salicylic acid response than Psm ES4326. Additional mutants were profiled using a custom array. Profiles of pbs3 and ndr1 revealed major effects of these mutations and allowed PBS3 and NDR1 to be placed between the EDS1/PAD4 node and the SA synthesis node in the defense network. Comparison of coi1, dde2, and jar1 profiles showed that many genes were affected by coi1, but very few were affected by dde2 or jar1. Profiles of coi1 plants infected with Psm ES4326 were very similar to those of wild-type plants infected with bacteria unable to produce the phytotoxin coronatine, indicating that essentially all COI1-dependent gene expression changes in this system are caused by coronatine. This experiment consists of three biological replicates. For each genotype, two leaves per plant were pooled from three pots to prepare total RNA.
2008-07-18 | E-GEOD-11009 | biostudies-arrayexpress
Project description:Investigation of whole genome gene expression AdnA in Pseudomonas fluorescens, an ortholog of FleQ in P. aeruginosa, regulates both motility and flagella-mediated attachment to various surfaces. A whole genome microarray determined the AdnA transcriptome by comparing the gene expression pattern of wild-type Pf0-1 to that of Pf0-2x (adnA-) in broth culture. In the absence of AdnA, expression of 92 genes was decreased, while 11 genes showed increased expression. Analysis of 16 of these genes fused to lacZ confirmed the microarray results. Several genes were further evaluated for their role in motility and biofilm formation. Two genes, Pfl01_1508 and Pfl01_1517, affected motility and had different effects on biofilm formation in Pf0-1. These two genes are predicted to encode proteins similar to the glycosyl transferases FgtA1 and FgtA2, which have been shown to be involved in virulence and motility in P. syringae. Three other genes, Pfl01_1516, Pfl01_1572, and Pfl01_1573, not previously associated with motility and biofilm formation in Pseudomonas had similar affects on biofilm formation in Pf0-1. Deletion of each of these genes led to different motility defects. Our data revealed an additional level of complexity in the control of flagella function beyond the core genes known to be required, and may yield insights into processes important for environmental persistence of P. fluorescens Pf0-1. Catalog design for Pseudomonas fluorescens Pf0-1 (Taxonomy Id: 205922) covering NC_007492. Probes selected for 5736/5736 sequences. Median number of probes/sequence is 13 with an average of 13.00. Each probe will be replicated 5 times on the chip. Probes are randomly distributed over the surface of the array. Unused features have been filled with randomly generated probes of comparable GC content.
2011-11-30 | E-GEOD-33865 | biostudies-arrayexpress
Project description:The purpose of this study was to investigate gene expression in SIDS cases and controls, in order to uncover genes that are differentially expressed in the two groups. Tissue from brain, heart and liver from 15 SIDS cases and 15 controls were included in this study, and mRNA expression was determined using the Illumina whole genome gene expression DASL HT assay WGDASL_HT ASSAY 15018210_D
2016-03-01 | E-GEOD-70422 | biostudies-arrayexpress
Project description:AtGenExpress: A multinational coordinated effort to uncover the transcriptome of the multicellular model organism Arabidopsis thaliana; The activity of genes and their encoded products can be regulated in several ways, but transcription is the primary level, since all other modes of regulation (RNA splicing, RNA and protein stability, etc.) are dependent on a gene being transcribed in the first place. The importance of transcriptional regulation has been underscored by the recent flood of global expression analyses, which have confirmed that transcriptional co-regulation of genes that act together is the norm, not the exception. Moreover, many studies suggest that evolutionary change is driven in large part by modifications of transcriptional programs. An essential first step toward deciphering the transcriptional code is to determine the expression pattern of all genes. With this goal in mind, an international effort to develop a gene expression atlas of Arabidopsis has been underway since fall 2003. This project, dubbed AtGenExpress, is funded by the DFG, and will provide the Arabidopsis community with access to a large set of Affymetrix microarray data. As part of this collaboration, we have generated expression data from 80 biologicaly different samples in triplicate. Half of a Columbia plant leaf was injected with Pseudomonas syringae. The other half of the leaf was collected on a time course for analysis. Experimenter name = Xinnian Dong; Experimenter institute = AtGenExpress Experiment Overall Design: 32 samples were used in this experiment
2008-06-14 | E-GEOD-5685 | biostudies-arrayexpress
Project description:We implemented transcriptional analysis methods using cDNA and high-throughput sequencing data to identify HrpL-regulated genes for six strains of Pseudomonas syringae Each Pseudomonas syringae strains was transformed with either pBAD::EV or pBAD containing native hrpL sequence. Strains were grown in MM media supplemented with arabinose and collected 1, 3, and 5 hours post arabinose treatment. RNA was extracted for each time point and mixed at a 1/3 ratio. After removal of rRNA, double stranded cDNA was generated and library prepared accordeing to Illumina protocols.
2014-01-01 | E-GEOD-46930 | biostudies-arrayexpress