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Integrative Genomics Positions MKRN1 as a Novel Ribonucleoprotein within the Embryonic Stem Cell Gene Regulatory Network

ABSTRACT: In this study we affinity purified FLAG-tagged MKRN1 from mouse embryonic stem cells constitutively expressing FLAG:MKRN1 that were pre-treated or untreated with the proteasome inhibitor MG132. FLAG:MKRN1 repeatedly co-immunoprecipitated with 48 proteins irrespective of MG132 treatment. Many of the MKRN1-associated proteins are well-characterized RNA-binding proteins, and post-translational regulators of gene expression.

OTHER RELATED OMICS DATASETS IN: PRJNA255236GSE59394PRJNA255235PRJNA255234

INSTRUMENT(S): LTQ

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Embryonic Stem Cell

SUBMITTER: Gareth Palidwor

LAB HEAD: Dr. William Stanford

PROVIDER: PXD001191 | Pride | 2018-10-19

REPOSITORIES: Pride

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Integrative genomics positions MKRN1 as a novel ribonucleoprotein within the embryonic stem cell gene regulatory network.

Cassar Paul A PA Carpenedo Richard L RL Samavarchi-Tehrani Payman P Olsen Jonathan B JB Park Chang Jun CJ Chang Wing Y WY Chen Zhaoyi Z Choey Chandarong C Delaney Sean S Guo Huishan H Guo Hongbo H Tanner R Matthew RM Perkins Theodore J TJ Tenenbaum Scott A SA Emili Andrew A Wrana Jeffrey L JL Gibbings Derrick D Stanford William L WL

EMBO reports 20150811 10

In embryonic stem cells (ESCs), gene regulatory networks (GRNs) coordinate gene expression to maintain ESC identity; however, the complete repertoire of factors regulating the ESC state is not fully understood. Our previous temporal microarray analysis of ESC commitment identified the E3 ubiquitin ligase protein Makorin-1 (MKRN1) as a potential novel component of the ESC GRN. Here, using multilayered systems-level analyses, we compiled a MKRN1-centered interactome in undifferentiated ESCs at the ...[more]

PMID: 26265008

Similar Datasets

Integrative Genomics Positions MKRN1 as a Novel Ribonucleoprotein within the Embryonic Stem Cell Gene Regulatory Network

Project description:For identification of proteins that associate with Makorin1 (MKRN1) in RNA-dependent and RNA-independent manners, we affinity purified FLAG-tagged Makorin1 (MKRN1) from mouse embryonic stem cells constitutively expressing FLAG:MKRN1. Anti-FLAG control immunoprecipitations were performed from a FLAG vectrol control (FLAG:Ctrl) mouse embryonic stem cell line that did not express FLAG:MKRN1. Following FLAG immunoprecipitation, anti-FLAG beads from FLAG:MKRN1 and FLAG:Ctrl immunoprecipitations were split into separate tubes such that half of the beads were digested with 200Ã∞â≈ Ã≠Âµg/mL RNase A while the other half of the beads were undigested. RNase A-digested and undigested immunoprecipitates were subjected to LC-MS/MS analysis. Of the 48 RNA-related proteins previously identified to associate with FLAG:MKRN1, L1TD1, PABPC1, PABPC4, YBX1, IGF2BP1 and UPF1 were found to remain associated with FLAG:MKRN1 in the presence of RNase A.

2015-07-21 | PXD001291 | Pride

Post-transcription initiation function of the ubiquitous SAGA complex in tissue-specific gene activation (ChIP-seq)

Project description:SAGA is a highly conserved transcriptional co-activator complex involved in multiple steps of transcription with activities that function both pre and post initiation. Loss of individual subunits results in developmental defects, suggesting a role in development. To better understand the roles of SAGA functions in developmental gene expression and it's relationship with RNA polymerase II, we examined its composition, binding profile, and the effects of subunit loss on gene expression in two distinct cell types in late stage Drosophila embryos: muscle and neurons. Chromatin IP of FLAG-tagged SAGA subunit Ada2b, and RNA polymerase II (antibody 4H8), was performed in neuronal (elav+) or muscle (mef2+) cells isolated by FACS from late stage embryos, and compared to whole cell extracts (input). The control constists of an IP performed in neuronal cells from a non-tagged strain.

2011-08-22 | E-GEOD-29513 | biostudies-arrayexpress

The RPB2 flap loop of human RNA polymerase II is dispensable for transcription initiation and elongation.

Project description:Deletion of the flap loop in the Rp2 subunit of human RNAPII did not alter the ability of human RNAPII to initiate transcription or elongate the initiated transcripts in vivo and in vitro, and was also dispensable for elongation factor-mediated enhancement and inhibition of transcript elongation in vitro. ChIP:chip of wild-type and flap deletion human RNAPII revealed that the mutant RNAPII was not defective for promoter binding, initiation and elongation in vivo. Eight ChIP datasets were generated as log2(IP/input) fluorescence intensities using anti-FLAG IP of wild-type RPB2-FLAG cells in biological triplicate, anti-FLAG IP of delta-FL RPB2 FLAG cells in biological triplicate, and anti-RPB1 CTD, 8WG16 of both wild-type and delta-FL cells in biological duplicate. The two 8WG16- IP datasets were combined to measure the distribution of RPB1 signal.

2011-06-24 | E-GEOD-29734 | biostudies-arrayexpress

Dissecting psa locus regulation in Yersinia pestis - ChIP-seq

Project description:We looked at binding sites in wild type Yersinia pestis using a strain with psaE either FLAG-tagged or His-tagged.

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ChIP-chip analysis of GlnR-FLAG binding sites in Streptomyces venezuelae under varying conditions of nitrogen availibility

Project description:ChIP-chip analysis of GlnR-FLAG binding sites in Streptomyces venezuelae under varying conditions of nitrogen availibility

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Selective mRNA sequestration by OLIGOURIDYLATE BINDING PROTEIN 1 contributes to translational control during hypoxia in Arabidopsis

Project description:Low oxygen stress dynamically regulates the translation of cellular mRNAs as a means of energy conservation in seedlings of Arabidopsis thaliana. Most of the highly hypoxia-induced mRNAs are recruited to polysomes and actively translated, whereas other cellular mRNAs become translationally inactive and are either targeted for stabilization or degradation. Here we identify the involvement of OLIGOURIDYLATE BINDING PROTEIN 1 (UBP1), a triple RNA Recognition Motif protein, in dynamic and reversible aggregation of translationally repressed mRNAs during hypoxia. Mutation or downregulation of UBP1C interferes with seedling establishment and reduces survival of low oxygen stress. By use of messenger ribonucleoprotein immunopurification, we show that UBP1C constitutively binds a subpopulation of mRNAs characterized by U-rich 3M-bM-^@M-^Y-untranslated regions under normoxic conditions. During hypoxia, UBP1C association with non-U-rich mRNAs is enhanced concomitant with its aggregation into microscopically visible cytoplasmic foci, referred to as UBP1 stress granules (SGs). This UBP1C-mRNA association occurs as global levels of protein synthesis decline. Upon reoxygenation, rapid UBP1 SG disaggregation coincides with the return of the stabilized mRNAs to polysomes. The mRNAs that are highly induced and translated during hypoxia largely circumvent UBP1C sequestration. Thus, UBP1 is established as a component of dynamically assembled cytoplasmic mRNPs that sequester mRNAs that are poorly translated during a transient low energy stress. Immunoprecipated RNA associated with Arabidopsis UBP1C (IP) was compared with total cellular RNA from light (L), mock dark (D), 2 h hypoxia, and 2 h hypoxia + 20 min reoxygenation treated samples with duplicate hybridizations to the Affymetrix ATH1 Genechip array.

2014-01-13 | E-GEOD-49411 | biostudies-arrayexpress

ArcA Chip-chip from Escherichia coli MG1655 K-12, WT and M-bM-^HM-^FarcA strains

Project description:Mapping the occupancy of ArcA throughout the genome of Escherchia coli MG1655 K-12 using an affinity purified antibody under anaerobic and aerobic growth conditions. As a control, we also performed ChIP-chip onArcA in a M-bM-^HM-^FarcA mutant strain of Escherchia coli MG1655 K-12. Described in the manuscript The response regulator ArcA uses a diverse binding site architechture to globally regulate carbon oxidation in E. coli Mapping of occupancy of ArcA in the genome of Escherchia coli MG1655 K-12 during anaerobic fermentation and aerobic respiration. Immunoprecipitated DNA compared to INPUT for each sample.

2013-10-17 | E-GEOD-46413 | biostudies-arrayexpress

Makorin 1 controls embryonic patterning by alleviating Bruno-mediated repression of oskar translation

Project description:Makorins are an evolutionary conserved family of proteins that contain C3H-type zinc finger modules and a RING E3 ubiquitin ligase domain. Previous analysis indicated a maternal role for Makorin 1 (Mkrn1) in Drosophila embryonic patterning and germ cell specification, but the underlying mechanism has remained elusive. Here, we show that Mkrn1 is specifically required for translational activation of oskar, which encodes a critical regulator of axis specification and germ plasm assembly. We demonstrate that Mkrn1 interacts with poly(A) binding protein (pAbp) and specifically binds osk 3’ UTR adjacent to A-rich sequences. The binding of Mkrn1 to osk 3’UTR occurs in a region that overlaps with Bruno responsive elements (BRE), previously shown to have a dual role in regulating osk translation. We observe an increased association of the translational repressor Bruno (Bru) with osk mRNA upon depletion of Mkrn1, implying that both proteins compete with each other for osk binding. Consistently, reducing Bru dosage is sufficient to partially rescue osk translation and the embryonic lethality associated with Mkrn1 alteration. Thus, we conclude that Mkrn1 controls embryonic patterning and germ cell formation by specifically activating osk translation via displacing Bru from its 3’ UTR

2020-01-29 | PXD011802 | Pride

The evolutionary impact of Intragenic FliA Promoters

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Identification of OsSGS3a-FLAG-interacting proteins by IP-MS/MS

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