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Analysis of E2F2 interacting proteins

ABSTRACT: In an attempt to identify proteins interacting with transcription factor E2F2, proteins extracted from HA-E2F2 overexpressing HEK293T cells were were subjected to immunoprecipitation (IP) with an anti-HA antibody or, as a negative control (NegCt), with an anti-T antibody. Immunoprecipitated proteins were eluted, digested with trypsin, and the resulting peptides were analyzed by LC-MS/MS. The whole procedure was repeated several times in an attempt to gauge reproducibility (nine replicates of the anti-HA IP and six replicates of the control IP).

INSTRUMENT(S): Q-Tof micro

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hek-293t Cell, Kidney Cell

SUBMITTER: Nerea Osinalde

LAB HEAD: Nerea Osinalde

PROVIDER: PXD000026 | Pride | 2013-01-18

REPOSITORIES: pride

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			Action	DRS
	NEREA_IP30HA1_080120_001.pkx	Other
	NEREA_IP30HA1_080120_001.raw.rar	Raw
	NEREA_IP30HA2_080120_001.pkx	Other
	NEREA_IP30HA2_080120_001.raw.rar	Raw
	NEREA_IP30HA3_080120_001.pkx	Other

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Publications

The nuclear protein ALY binds to and modulates the activity of transcription factor E2F2.

Osinalde Nerea N Olea Miguel M Mitxelena Jone J Aloria Kerman K Rodriguez Jose Antonio JA Fullaondo Asier A Arizmendi Jesus M JM Zubiaga Ana M AM

Molecular & cellular proteomics : MCP 20130107 5

E2F transcription factors control the expression of genes involved in a variety of essential cellular processes and consequently their activity needs to be tightly regulated. Protein-protein interactions are thought to be key modulators of E2F activity. To gain insight into the mechanisms that regulate the activity of E2F2, we searched for novel proteins that associate with this transcription factor. We show that the nuclear protein ALY (THO complex 4), originally described as a transcriptional ...[more]

PMID: 23297349

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Analysis of E2F2 interacting proteins

Project description:In an attempt to identify proteins interacting with transcription factor E2F2, proteins extracted from HA-E2F2 overexpressing HEK293T cells were were subjected to immunoprecipitation (IP) with an anti-HA antibody or, as a negative control (NegCt), with an anti-T antibody. Immunoprecipitated proteins were eluted, digested with trypsin, and the resulting peptides were analyzed by LC-MS/MS. The whole procedure was repeated several times in an attempt to gauge reproducibility (nine replicates of the anti-HA IP and six replicates of the control IP).

2016-03-09 | MSV000079567 | MassIVE

Adaptor candidates

Project description:IP of candidate adaptor Ada5/Gdi1, tagged with HA, versus Mock (untagged) replicates. IP associated RNA was assayed. A replicate experimental design type is where a series of replicates are performed to evaluate reproducibility or as a pilot study to determine the appropriate number of replicates for a subsequent experiments. GeneticModification: HA-tagged candidate gene or control

2012-11-15 | GSE33747 | GEO

TIAR represses c-Myc biosynthesis following ultraviolet light irradiation

Project description:RNA was isolated from material that had been subjected to immunoprecipitation (IP) from RKO cells that were either left untreated or irradiated with 20 J/m2 UVC and collected 1h later; IP assays were carried out using either an antibody recognizing RNA-binding protein TIAR, or using a control IgG1 antibody. RNA was reverse-transcribed in the presence of [alpha-33P]dCTP and the radiolabeled product used to hybridize human cDNA arrays. The experiment was repeated using three independent sample sets. The samples were numbered Tc-1, Tc-2, Tc-3, Tuv-1, Tuv-2, Tuv-3, IgG1c-1, IgG1c-2, IgG1c-3, IgG1uv-1, IgG1uv-2, and IgG1uv-3. ‘Tc’ denotes RNA from IP reactions using untreated cells and anti-TIAR antibody, ‘Tuv’ denotes RNA from IP reactions using UVC-treated cells and anti-TIAR antibody, ‘IgG1c’ denotes RNA from IP reactions using untreated cells and anti-IgG1 antibody, and ‘IgG1uv’ denotes RNA from IP reactions using UVC-treated cells and anti-IgG1 antibody. The numbers 1, 2 and 3 correspond to the three independent experimental datasets. Keywords = post-transcriptional / translation / nascent protein synthesis / stress response / ribonomics Keywords: ordered

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Concurrent Versus Individual Binding of HuR and AUF1 to Common Labile Target mRNAs

Project description:RNA was isolated from material that had been immunoprecipitated (IP) from Hela cells using antibodies recognizing RNA-binding proteins HuR or AUF1, as well as using a control IgG1 antibody. RNA was reverse-transcribed in the presence of [alpha-33P]dCTP and the radiolabeled product used to hybridize human cDNA arrays. The experiment was repeated using three independent sample sets. The samples were numbered HuR-1, HuR-2, HuR-3, AUF1-1, AUF1-2, AUF1-3, IgG1-1, IgG1-2, IgG1-3. HuR represents RNA from IP reactions using an anti-HuR antibody, AUF1 represents RNA from IP reactions using an anti-AUF1 antibody and, IgG1 represents RNA from IP reactions using an anti-IgG1 antibody. The numbers 1, 2 and 3 correspond to the three independent experimental datasets. Keywords = RNa-binding protein Keywords = mRNA stability Keywords = exosome Keywords = polysome Keywords = RNA motif Keywords: ordered

2004-05-04 | GSE1361 | GEO

DiazMeco_Increased translation driven by a non-canonical EZH2 cistrome creates a synthetic vulnerability in enzalutamide-resistant prostate cancer_set1

Project description:Identification of interacting partners from immuno-precipitated HA-EZH2 samples. Endogenous Co-IP was performed using 1.5 mg of nuclear lysate with anti-HA antibody after transfecting HEK 293T cells with HA-EZH2 plasmid. Two IP-ed samples were submitted to the core for analysis, HEK293T-sgC and sgPRKCI.

2024-05-07 | MSV000094704 | MassIVE

TIAR represses c-Myc biosynthesis following ultraviolet light irradiation

Project description:RNA was isolated from material that had been subjected to immunoprecipitation (IP) from RKO cells that were either left untreated or irradiated with 20 J/m2 UVC and collected 1h later; IP assays were carried out using either an antibody recognizing RNA-binding protein TIAR, or using a control IgG1 antibody. RNA was reverse-transcribed in the presence of [alpha-33P]dCTP and the radiolabeled product used to hybridize human cDNA arrays. The experiment was repeated using three independent sample sets. The samples were numbered Tc-1, Tc-2, Tc-3, Tuv-1, Tuv-2, Tuv-3, IgG1c-1, IgG1c-2, IgG1c-3, IgG1uv-1, IgG1uv-2, and IgG1uv-3. âTcâ denotes RNA from IP reactions using untreated cells and anti-TIAR antibody, âTuvâ denotes RNA from IP reactions using UVC-treated cells and anti-TIAR antibody, âIgG1câ denotes RNA from IP reactions using untreated cells and anti-IgG1 antibody, and âIgG1uvâ denotes RNA from IP reactions using UVC-treated cells and anti-IgG1 antibody. The numbers 1, 2 and 3 correspond to the three independent experimental datasets. Keywords = post-transcriptional / translation / nascent protein synthesis / stress response / ribonomics Keywords: ordered

2010-06-09 | E-GEOD-1433 | biostudies-arrayexpress

Concurrent Versus Individual Binding of HuR and AUF1 to Common Labile Target mRNAs

2010-06-09 | E-GEOD-1361 | biostudies-arrayexpress

Human 293T -cGAS-MYO1F-KAT2A,LC-MS

Project description:To identify cGAS acetylation sites, 293T cells coexpressing the PCDH-EGFP-puro empty vector or PCDH-EGFP-puro-Flag-cGAS with KAT2A and MYO1F were subjected to immunoprecipitation with an anti-Flag antibody. After washing four times with IP buffer, the protein was eluted with IP buffer containing 0.1% SDS and analyzed by mass spectrometry.

2024-12-14 | PXD057694 | Pride

Identification of the BRD1 interactome and its impact on schizophrenia risk

Project description:We identified protein-protein interactions and chromatin binding sites for two isforms of BRD1 (BRD1-S and BRD1-L) using Co-IP MS/MS and ChIP-seq. BRD1 isoforms were cloned, epitope-tagged (pcDNA 6 V5-His6, Lifetechnologies) and stably expressed in HEK293T cells. Chromatin immunoprecipitations were performed using anti V5-antibody conjugated agarose beads (Sigma-Aldrich) and anti HA antibody conjugated agarose beads (Sigma-Aldrich) as controls

2015-10-29 | E-GEOD-62811 | biostudies-arrayexpress

Neuron-specific RNA responses to EAE and cuprizone

Project description:Ribotag mice were crossed to synapsin-CRE mice to induce expression of HA-tagged Rpl22 subunits in neurons. Mice were fed cuprizone to induce demyelination or were induced with MOG(35-55) peptide to establish experimental autoimmune encephalomyelitis. Brain was collected from EAE mice at 18 days after induction (peak disease) and from cuprizone-fed mice at 6 weeks on chow. Brain was also collected from unmanipulated control mice fed normal chow. Brain lysates were immunoprecipated with anti-HA antibody to isolated neuron-specific polyribosomes. RNA was extracted from the IP and analyzed by Affymetrix GeneChip® Mouse Gene 2.0 ST Arrays.

2023-09-05 | GSE241781 | GEO

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