Cell‐line specific chromatin acetylation at the Sox10–Pax3 enhancer site modulates the <i>RET</i> proto‐oncogene expression

Puppo, Francesca; Griseri, Paola; Fanelli, Mirco; Schena, Francesca; Romeo, Giovanni; Pelicci, PierGiuseppe; Ceccherini, Isabella; Ravazzolo, Roberto; Patrone, Giovanna

doi:10.1016/s0014-5793(02)02957-5

Cited by 19 publications

(22 citation statements)

References 27 publications

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“…At promoter region, the main driver of RET activation appears to be the increase of H3K4me3 levels while no significant changes of the methylation state of H3K27 and H3K9 were observed. Acetylation of histone H3 increased at both enhancer and promoter regions in concomitance with RET activation, which is in agreement with previous reports (20). At RET enhancer region we observed a simultaneous presence of histone modifications associated with transcriptional repression (H3K27me3) and activation (H3K4me2) (Figure 3).…”

Section: Discussionsupporting

confidence: 93%

Chromatin and DNA methylation dynamics during retinoic acid-induced RET gene transcriptional activation in neuroblastoma cells

Angrisano¹,

Sacchetti²,

Natale³

et al. 2010

Nucleic Acids Research

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Although it is well known that RET gene is strongly activated by retinoic acid (RA) in neuroblastoma cells, the mechanisms underlying such activation are still poorly understood. Here we show that a complex series of molecular events, that include modifications of both chromatin and DNA methylation state, accompany RA-mediated RET activation. Our results indicate that the primary epigenetic determinants of RA-induced RET activation differ between enhancer and promoter regions. At promoter region, the main mark of RET activation was the increase of H3K4me3 levels while no significant changes of the methylation state of H3K27 and H3K9 were observed. At RET enhancer region a bipartite chromatin domain was detected in unstimulated cells and a prompt demethylation of H3K27me3 marked RET gene activation upon RA exposure. Moreover, ChIP experiments demonstrated that EZH2 and MeCP2 repressor complexes were associated to the heavily methylated enhancer region in the absence of RA while both complexes were displaced during RA stimulation. Finally, our data show that a demethylation of a specific CpG site at the enhancer region could favor the displacement of MeCP2 from the heavily methylated RET enhancer region providing a novel potential mechanism for transcriptional regulation of methylated RA-regulated loci.

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Section: Discussionsupporting

confidence: 93%

Chromatin and DNA methylation dynamics during retinoic acid-induced RET gene transcriptional activation in neuroblastoma cells

Angrisano¹,

Sacchetti²,

Natale³

et al. 2010

Nucleic Acids Research

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“…27 In a previous paper, we demonstrated that the histone deacetylase inhibitor NaB increased RET transcription in cells displaying low mRNA levels while it had no effect in cells already expressing high mRNA levels. 22 Moreover, we identified two RET upstream regions sensitive to NaB treatment, the SOX10/ Pax3 enhancer and the minimal promoter, supporting the view that acetylation of RET histones is a crucial and physiological regulatory mechanism of RET transcription. 22 Here we have shown that NaB treatment induces the appearance of RET transcript in RET negative cells, such as lymphoblasts and the SK-N-MC cell line.…”

Section: Discussionsupporting

confidence: 71%

“…22 Moreover, we identified two RET upstream regions sensitive to NaB treatment, the SOX10/ Pax3 enhancer and the minimal promoter, supporting the view that acetylation of RET histones is a crucial and physiological regulatory mechanism of RET transcription. 22 Here we have shown that NaB treatment induces the appearance of RET transcript in RET negative cells, such as lymphoblasts and the SK-N-MC cell line. Starting from such an observation we have developed a novel and useful approach to analyse the RET transcription product.…”

Section: Discussionsupporting

confidence: 71%

“…22 In particular, NaB treatment increased RET transcription in IMR32 cells, already displaying low levels of its mRNA, while it had no effect on MTC-TT cells, characterised by a plateau level of RET mRNA. 22 Accordingly, we also demonstrated that NaB treatment increased the endogenous level of RET histones acetylation (generally associated with transcriptionally active chromatin) in IMR32 but not in MTC-TT cells. These data strongly suggest that NaB treatment mimics a physiological regulatory mechanism of the RET gene.…”

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confidence: 87%

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Rescue of human RET gene expression by sodium butyrate: a novel powerful tool for molecular studies in Hirschsprung disease

Griseri

Patrone

Puppo

et al. 2003

Gut

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Background:The RET gene encodes a tyrosine kinase receptor involved in different human neurocristopathies, such as specific neuroendocrine tumours and Hirschsprung disease (HSCR). Gene expression is developmentally regulated and the RET transcript is undetectable in most adult cells, including lymphocytes. The impossibility of performing functional studies on RET mRNA has to date limited the detection and characterisation of an indefinite proportion of gene anomalies that cannot be identified by conventional DNA genomic screening in HSCR cases. Aims: Development of a protocol suitable to activate RET expression in RET negative cell lines and therefore to investigate directly RET mRNA, extending the conventional gene mutation analysis to detection of splicing anomalies and impaired expression of the RET gene. Methods: The effect of sodium butyrate (NaB), a histone deacetylase inhibitor, on rescuing RET expression was tested by one round of reverse transcription-polymerase chain reaction from total RNA of treated lymphoblasts from both HSCR patients and control individuals. Results: Analysis of RET expression was possible by NaB treatment of RET negative cells, such as lymphoblasts. This treatment allowed us to detect impaired RET expression as well as a splicing defect in two HSCR patients previously believed to be devoid of any gene abnormality. Conclusions:The full application of the proposed protocol in most of the unexplained HSCR cases will allow us to establish the precise role of RET not only in causing but also in predisposing to HSCR pathogenesis.T he RET proto-oncogene (REarranged during Transfection) encodes a tyrosine kinase receptor predominantly expressed during neural crest development and involved in different human neurocristopathies.1 Germline RET mutations have been found in association with both inherited cancer syndromes, including multiple endocrine neoplasia type 2A (MEN2A), type 2B (MEN2B), and familial medullary thyroid carcinoma, and Hirschsprung disease (HSCR), a developmental disorder characterised by the absence of enteric neurones in variable lengths of the distal gastrointestinal tract.2 Loss of function, low penetrant RET mutations have been identified in only 10-40% of total HSCR cases while a small proportion of patients (5-10%) show alterations in other genes, generally related to the developmental programme of neural crest cells.3 4 By using sibpair analysis, a recent study has recognised three HSCR susceptibility loci at 3p21, 10q11, and 19q12 which may clarify the molecular basis of a great proportion of unexplained HSCR cases. 5 The locus in 10q11 probably corresponds to RET, confirming the central role played by this gene in HSCR pathogenesis. Taking into account the persistent failure to detect RET coding sequence mutations in a proportion of familial cases found to be linked to the gene, it has recently been suggested that genetic defects in introns and in the promoter region can contribute in some cases to the HSCR phenotype. [5][6][7] Moreover, several common polymorphi...

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“…RET is repressed in most tissues and cells, and its expression is associated with a transition from a repressive chromatin to an open, permissive chromatin [35], [36]. It is plausible to speculate that association of HOXB5 with HOX binding sequence and interaction with NKX2-1 at the transcription start site, may induce conformational change of the RET promoter bringing transcription factors in close proximity of minimal promoter facilitating the recruitment of basal transcription factors mediating RET transcription.…”

Section: Discussionmentioning

confidence: 99%

HOXB5 Cooperates with NKX2-1 in the Transcription of Human RET

et al. 2011

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The enteric nervous system (ENS) regulates peristaltic movement of the gut, and abnormal ENS causes Hirschsprung's disease (HSCR) in newborns. HSCR is a congenital complex genetic disorder characterised by a lack of enteric ganglia along a variable length of the intestine. The receptor tyrosine kinase gene (RET) is the major HSCR gene and its expression is crucial for ENS development. We have previously reported that (i) HOXB5 transcription factor mediates RET expression, and (ii) mouse with defective HOXB5 activity develop HSCR phenotype. In this study, we (i) elucidate the underlying mechanisms that HOXB5 mediate RET expression, and (ii) examine the interactions between HOXB5 and other transcription factors implicated in RET expression. We show that human HOXB5 binds to the promoter region 5′ upstream of the binding site of NKX2-1 and regulates RET expression. HOXB5 and NKX2-1 form a protein complex and mediate RET expression in a synergistic manner. HSCR associated SNPs at the NKX2-1 binding site (-5G>A rs10900296; -1A>C rs10900297), which reduce NKX2-1 binding, abolish the synergistic trans-activation of RET by HOXB5 and NKX2-1. In contrast to the synergistic activation of RET with NKX2-1, HOXB5 cooperates in an additive manner with SOX10, PAX3 and PHOX2B in trans-activation of RET promoter. Taken together, our data suggests that HOXB5 in coordination with other transcription factors mediates RET expression. Therefore, defects in cis- or trans-regulation of RET by HOXB5 could lead to reduction of RET expression and contribute to the manifestation of the HSCR phenotype.

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Cell‐line specific chromatin acetylation at the Sox10–Pax3 enhancer site modulates the RET proto‐oncogene expression

Cited by 19 publications

References 27 publications

Chromatin and DNA methylation dynamics during retinoic acid-induced RET gene transcriptional activation in neuroblastoma cells

Chromatin and DNA methylation dynamics during retinoic acid-induced RET gene transcriptional activation in neuroblastoma cells

Rescue of human RET gene expression by sodium butyrate: a novel powerful tool for molecular studies in Hirschsprung disease

HOXB5 Cooperates with NKX2-1 in the Transcription of Human RET

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