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Gene regulation by phosphomimetic Myc (MycD), cytosine arabinoside (AraC) and retinoic acid (RA)

ABSTRACT: We aimed to investigate whether the previously observed synergistic effects of AraC and RA in stimulating leukemia differentiation could be at least partially dependent on Pak2-mediated phosphorylation of Myc. Genome-wide analysis of NB4 cells treated with RA, AraC with RA, or MycD with RA revealed a significant 60% overlap between RA-target genes superactivated by AraC with RA, or MycD with RA, with respect to RA alone. NB4 cells infected with ER-tagged inducible empty vector (CTL), MycD or MycA were treated with 4-OHT (200 nM, 16 h) to induce MycD-ER and MycA-ER expression. CTL cells were either vehicle or AraC (25 μM) treated for 16 h. Short treatment of RA (10 nM, 4 h) was performed in CTL, AraC and MycD-expressing cells. Four biologically independent experiments were performed at each time.

ORGANISM(S): Homo sapiens

SUBMITTER: Luciano Di Croce

PROVIDER: E-GEOD-29213 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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E-box-independent regulation of transcription and differentiation by MYC.

Uribesalgo Iris I Buschbeck Marcus M Gutiérrez Arantxa A Teichmann Sophia S Demajo Santiago S Kuebler Bernd B Nomdedéu Josep F JF Martín-Caballero Juan J Roma Guglielmo G Benitah Salvador Aznar SA Di Croce Luciano L

Nature cell biology 20111023 12

MYC proto-oncogene is a key player in cell homeostasis that is commonly deregulated in human carcinogenesis(1). MYC can either activate or repress target genes by forming a complex with MAX (ref. 2). MYC also exerts MAX-independent functions that are not yet fully characterized(3). Cells possess an intrinsic pathway that can abrogate MYC-MAX dimerization and E-box interaction, by inducing phosphorylation of MYC in a PAK2-dependent manner at three residues located in its helix-loop-helix domain(4 ...[more]

PMID: 22020439

Similar Datasets

Gene regulation by phosphomimetic Myc (MycD), cytosine arabinoside (AraC) and retinoic acid (RA)

Project description:We aimed to investigate whether the previously observed synergistic effects of AraC and RA in stimulating leukemia differentiation could be at least partially dependent on Pak2-mediated phosphorylation of Myc. Genome-wide analysis of NB4 cells treated with RA, AraC with RA, or MycD with RA revealed a significant 60% overlap between RA-target genes superactivated by AraC with RA, or MycD with RA, with respect to RA alone.

2012-01-05 | GSE29213 | GEO

Gene regulation by phosphomimetic Myc and retinoic acid

Project description:We aimed to investigate the E-box and Max-independent functions of the oncogene Myc in gene regulation and differentiation of leukemic cells. Expression in HL60 leukemic cells of a mutant form of Myc with impaired Max and E-box interaction by replacing the phosphorylation sites of Pak2 kinase to aspartic acid (MycD), resulted in downregulation of 235 genes and, interestingly, upregulation of 586 genes. Eleven percent of the genes upregulated by MycD coincided with those stimulated by a short treatment of retinoic acid (RA) alone. When leukemic cells expressing MycD were stimulated with RA, the overlap with the RA-alone signature increased to 32% (492/1556 genes). Importantly, 320 of these 492 bona-fide direct RA target genes were specifically superactivated in presence of MycD (MycD+RA>RA), and not by MycA (MycD+RA>MycA+RA), further suggesting a synergy between the two pathways. The list of superactivated genes included important regulators of development and differentiation such as GATA6, ICAM1 and HOX genes, whose expression was validated in independent experiments by RT-qPCR. Induced gene expression in HL60 cells was measured after a short-treatment of retinoic acid (100nM, 4 hours). Cells were either wild-type or expressing ER-tagged MycD or MycA. Four biologically independent experiments were performed at each time.

2012-01-05 | E-GEOD-24731 | biostudies-arrayexpress

Effects of CEBPA isoform expression on NB4 cells

Project description:Here we investigated the effects of CEBPA transcription factor expression on myeloid NB4 cells. The sequence of rat CEBPA was C-terminally fused to a promiscuous biotin ligase tag (BirA*) and NB4 cell lines were engineered to express the fusion protein under the control of a doxycycline inducible promoter. Three different NB4 cell lines were investigated that expressed (i) BirA* tag alone (ii) full length CEBPA isoform (P42) fused to BirA* (iii) truncated CEBPA isoform (P30) fused to BirA*. Cells were seeded in media supplemented with or without doxycycline.

2021-06-26 | E-MTAB-9947 | biostudies-arrayexpress

14028s araC RNA-Seq

Project description:14028s araC RNA-Seq

| PRJEB4670 | ENA

Salmonella 14028s AraC ChIP-Seq

Project description:We mapped the genome-wide binding of C-terminally FLAG-tagged AraC in S. enterica subsp. enterica serovar Typhimurium strain 14028s using ChIP coupled with deep sequencing (ChIP-seq). We identified five putative target loci for AraC: upstream of araB/araC, araE, araJ, STM14_0178, and within sseD.

2013-10-09 | E-MTAB-1915 | biostudies-arrayexpress

Transcription profiling of human ATRA treated NB4 cells

Project description:NB4 is an APL derived cell line, carrying the t(15;17) translocation and expressing the PML/RARa fusion protein. Still, an important question that remains to be addressed is whether PML/RARa target genes are transcriptionally suppressed in primary APL cells and re-activated in all-trans retinoic acid (ATRA) treated NB4 cells. Gene expression of NB4 cells treated with ATRA at different time points were analyzed. Experiment Overall Design: 6 samples at various times. No replicats.

2009-12-14 | E-GEOD-19201 | biostudies-arrayexpress

MG1655 araC arabinose transcription profiling

Project description:AraC is an Escherichia coli transcription factor that regulates genes in response to the presence/absence of arabinose. We used transcription profiling to determine RNA levels in Escherichia coli K-12 strain MG1655 and MG1655 delta araC grown either in the absence or the presence of arabinose. Thus, we identified known and novel AraC-regulated genes. Data presented here are raw .CEL files as well as fully analysed data using GeneSpring

2013-10-19 | E-MTAB-1916 | biostudies-arrayexpress

Gene expression profiling of ATRA-differentiated wild-type and TG2 knockout NB4 cells

Project description:Treatment of acute promyelocytic leukemia (APL) with all-trans-retinoic acid (ATRA) results in terminal differentiation of leukemic cells toward neutrophil granulocytes. Administration of ATRA leads to massive changes in gene expression, including down-regulation of cell proliferation-related genes and induction of genes involved in immune function. One of the most induced genes in APL NB4 cells is transglutaminase 2 (TG2). RNAi-mediated stable silencing of TG2 in NB4 cells (TG2-KD NB4) coupled with whole genome microarray analysis revealed that TG2 is involved in the expression of a large number of ATRA-regulated genes. The affected genes participate in granulocyte functions and their silencing lead to reduced adhesive, migratory and phagocytic capacity of neutrophils and less superoxide production. The expression of genes related to cell cycle control also changed, suggesting that TG2 regulates myeloid cell differentiation. CC chemokines CCL2, 3, 22, 24 and cytokines IL1B and IL8 involved in the development of differentiation syndrome (DS) are expressed at significantly lower levels in TG2-KD NB4 cells than in wild-type NB4 cells upon ATRA treatment. Based on our results, we propose that reduced expression of TG2 in differentiating APL cells may suppress effector functions of neutrophil granulocytes and attenuate the ATRA-induced inflammatory phenotype of DS. We used microarrays to detail the global program of gene expression underlying ATRA-induced differentiation of TG2 knockout NB4 cells. TG2 knockout NB4 cells were differentiated for 48 and 72 hours in the presence of ATRA and their gene expression profiles were compared to the wild-type cells at the same time points. Undifferentiated wild-type and TG2 knockout NB4 cells were used as untreated controls. Three biological replicates each.

2010-12-01 | E-GEOD-23702 | biostudies-arrayexpress

14028s araC RNA-Seq

Project description:We used RNA-seq to determine the effects of AraC and arabinose on RNA levels genome-wide in S. enterica. Wild-type or delta araC mutant cells were grown in both the presence and absence of 0.2% L-arabinose. RNA-Seq libraries from two independent biological replicate samples were sequenced for (i) wild-type cells with no treatment, (ii) wild-type cells treated with arabinose, (iii) delta araC cells with no treatment, (iv) delta araC cells treated with arabinose. Comparisons were made between (i) and (ii) [analysis file name: Salmonella RNA-seq processed WT without arabinose vs WT with arabinose.xls], between (i) and (iii) [analysis file name: Salmonella RNA-Seq proessed no arabinose WT vs no arabinose delta araC.xls], and between (ii) and (iv) [analysis file name: Salmonella RNA-Seq processed arabinose WT vs arabinose delta araC.xls]. All analysis files are available from https://www.ebi.ac.uk/arrayexpress/files/E-MTAB-1901/E-MTAB-1901.additional.1.zip

2013-10-04 | E-MTAB-1901 | biostudies-arrayexpress

RNA-sequencing analysis of NB4 cells overexpressing miR-125b

Project description:To better understand the mechanisms of blockage of myeloid differentiation and apoptosis and induction of proliferation by miR-125b, we proceeded to identify miR-125b target genes involved in these pathways. We analyzed the total cellular gene expression pattern by RNA-sequencing of the parental NB4 myeloid cell line and that transiently transfected with miR-125b. We generated four cDNA libraries corresponding to duplicates of miR-125b and control cells. Compare the gene expression levels in miR control transfected cells with that in miR-125b transfected NB4 cells.

2012-04-06 | E-GEOD-37061 | biostudies-arrayexpress

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