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LC-MS/MS after anti-PUM1 and anti-PUM2 co-immunoprecipitation.

ABSTRACT: In order to identify the protein co-factors of PUMILIO1 and PUMILIO2 in TCam-2 cell line, co-IP of these proteins performed followed mass spectrometry analysis for protein identification. Six biological replicates of co-IP with anti-PUM1, anti-PUM2 antibodies and anti-IgG were performed. Three biological replicates were performed without RNase A treatment, and another three biological replicates of co-IPs with 100 mg/ml RNase A.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Male Germ Cell, Cell Culture

SUBMITTER: Erkut Ilaslan

LAB HEAD: Jadwiga Jaruzelska

PROVIDER: PXD011948 | Pride | 2020-04-23

REPOSITORIES: pride

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Dataset's files

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			Action	DRS
	IGG-1.mgf	Mgf
	IGG-1.mzid.gz	Mzid
	IGG-1.pride.mztab.gz	Mztab
	IGG-1.raw	Raw
	IGG-2.mgf	Mgf

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Publications

Characterization of RNP Networks of PUM1 and PUM2 Post-Transcriptional Regulators in TCam-2 Cells, a Human Male Germ Cell Model.

Smialek Maciej J MJ Ilaslan Erkut E Sajek Marcin P MP Swiercz Aleksandra A Janecki Damian M DM Kusz-Zamelczyk Kamila K Wozniak Tomasz T Kotecki Maciej M Handschuh Luiza L Figlerowicz Marek M Jaruzelska Jadwiga J Jaruzelska Jadwiga J

Cells 20200416 4

Mammalian Pumilio (PUM) proteins are sequence-specific, RNA-binding proteins (RBPs) with wide-ranging roles. They are involved in germ cell development, which has functional implications in development and fertility. Although human PUM1 and PUM2 are closely related to each other and recognize the same RNA binding motif, there is some evidence for functional diversity. To address that problem, first we used RIP-Seq and RNA-Seq approaches, and identified mRNA pools regulated by PUM1 and PUM2 prote ...[more]

PMID: 32316190

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Project description:In human somatic cells, ZC3H11A has been recently characterized as an RNA-export protein that functions through its interaction with TREX complex proteins. In order to identify its interacting partners in embryonic cells, and to investigate if ZC3H11A maintains its association with the TREX complex in mouse embryonic stem cells (mESCs), we have performed co-immunoprecipitations (co-IPs) using anti-ZC3H11A, anti-THOC2 and anti-IgG antibodies followed by mass spectrometry (MS) analyses. Statistical analyses of detected MS intensities from the biological replicates (n=4) have revealed a number of proteins with statistically significant interaction with ZC3H11A and THOC2. Results in xlsx format contain the log-fold change in protein intensities in the ZC3H11A co-IP relative to the IgG co-IP and is presented along with the adjusted P-values.

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SMAD3 ChIP-on-chip on A549 cell line

Project description:The aim of this study is to identify SMAD3 binding targets affected by the TGFb1/SMAD3 signal transduction on a genome-wide scale Formaldehyde cross-linked, sonicated chromatin was prepared from non-stimulated A549 cell line and stimulated with TGFb1. Chromatin immunoprecipated with anti-SMAD3 antibody is labeled with Cy5 and mock IP (with anti-FLAG antibody) is labeled with Cy3 and co-hybridized on Agilent Human Promoter Set arrays. Two biological replicates were performed.

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