Characterization of BHC80 in BRAF–HDAC complex, involved in neuron-specific gene repression

Iwase, Shigeki; Januma, Aya; Miyamoto, Kiyoko; Shono, Naomi; Honda, Arata; Yanagisawa, Junn; Baba, Tadashi

doi:10.1016/j.bbrc.2004.07.163

Cited by 51 publications

(58 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The LSD1/Aof2-Ring1B/Rnf2 complex(es) remain to be characterized fully, but preliminary evidence suggests it is a new entity not identified previously perhaps because it contains a relatively small fraction of Ring1B/Rnf2 (and most likely LSD1/ Aof2 too) in the cell. In fact, this histone demethylase has been identified in a variety of subsets of the Braf-HDAC (BHC) repressing complexes (55)(56)(57)(58), but our analysis of Ring1B/ Rnf2 complexes did not detect either Braf35 or BHC80, two of the components of core BHC complexes (55,58). Thus, our data suggest that the LSD1/Aof2-Ring1B/Rnf2 complex is a novel complex not related to BHC.…”

Section: Discussioncontrasting

confidence: 55%

Proteomics Analysis of Ring1B/Rnf2 Interactors Identifies a Novel Complex with the Fbxl10/Jhdm1B Histone Demethylase and the Bcl6 Interacting Corepressor

Sánchez

Demmers³

et al. 2007

Molecular & Cellular Proteomics

208

192

View full text Add to dashboard Cite

Ring1B/Rnf2 is a RING finger protein member of the Polycomb group (PcG) of proteins, which form chromatinmodifying complexes essential for embryonic development and stem cell renewal and which are commonly deregulated in cancer. Ring1B/Rnf2 is a ubiquitin E3 ligase that catalyzes the monoubiquitylation of the histone H2A, one of the histone modifications needed for the transcriptional repression activity of the PcG of proteins. Ring1B/Rnf2 was shown to be part of two complexes, the PRC1 PcG complex and the E2F6.com-1 complex, which also contains non-PcG members, thus raising the prospect for additional Ring1B/Rnf2 partners and functions extending beyond the PcG. Here we used a high throughput proteomics approach based on the single step purification, using streptavidin beads, of in vivo biotinylated Ring1B/Rnf2 and associated proteins from a nuclear extract from erythroid cells and their identification by mass spectrometry. About 50 proteins were confidently identified of which 20 had not been identified previously as subunits of Ring1B/Rnf2 complexes. We found that histone demethylases LSD1/Aof2 and Fbxl10/Jhdm1B, casein kinase subunits, and the BcoR corepressor were among the new interactors identified. We also isolated an Fbxl10/Jhdm1B complex by biotinylation tagging to identify shared interacting partners with Ring1B/Rnf2. In this way we identified a novel Ring1B-Fbxl10 complex that also includes Bcl6 corepressor (BcoR), CK2␣, Skp1, and Nspc1/Pcgf1. The putative enzymatic activities and protein interaction and chromatin binding motifs present in this novel Ring1B-Fbxl10 complex potentially provide additional mechanisms for chromatin modification/recruitment to chromatin and more evidence for Ring1B/Rnf2 activities beyond those typically associated with PcG function. Lastly this work demonstrates the utility of biotinylation tagging for the rapid characterization of complex mixtures of multiprotein complexes achieved through the iterative use of this simple yet high throughput proteomics approach.Molecular & Cellular Proteomics 6:820 -834, 2007.In multicellular organisms, cell identity is controlled, at least in part, by epigenetic events, including DNA methylation and post-translational modifications of histones that lead to chromatin structure regulation (1). These modifications are carried out by protein complexes recruited through DNA sequences and/or specific recognition of modified histones. The Polycomb group (PcG) 1 of proteins, first identified genetically as regulators of Hox genes in the fly Drosophila melanogaster (for a review, see Ref. 2), is an example of such a complex. PcG functions cover many aspects of vertebrate development and tissue homeostasis by preventing the inappropriate activation of many transcription factor-coding genes and other genes involved in cell signaling and cell proliferation (3-7). Currently it is believed that PcG proteins play a role in setting the balance between proliferation and differentiation in normal development. Deregulation of PcG proteins disrupts such a ba...

show abstract

Section: Discussioncontrasting

confidence: 55%

Proteomics Analysis of Ring1B/Rnf2 Interactors Identifies a Novel Complex with the Fbxl10/Jhdm1B Histone Demethylase and the Bcl6 Interacting Corepressor

Sánchez

Demmers³

et al. 2007

Molecular & Cellular Proteomics

208

192

View full text Add to dashboard Cite

show abstract

“…BHC80 is differentially expressed in various cells and tissues (high in brain, low in many peripheral organs). Its role was still debated (Hakimi et al, 2002;Iwase et al, 2004;Shi et al, 2005;Lan et al, 2007). Our data, obtained with defective PC12 transfected or not with BHC80, demonstrate that association with BHC80 modulates negatively the repressor activity of REST and can thus contribute to the recovery of the neurosecretory competence of the cells.…”

Section: Introductionmentioning

confidence: 66%

“…The IgG2a ratspecific chromograninB (ChgB) monoclonal antibody (mAb; generated in the Milan laboratory) was as in the study by Borgonovo et al (2002); the anti-BHC80 polyclonal antibody (pAb; generated in the Ibaraki laboratory), as in the study by Iwase et al (2004). The anti-ICA512 mAb, the anti-secretogranin2 (Scg2) pAb, and the anti-uPAR pAb were the gifts of M. Solimena (University of Dresden, Dresden, Germany), P. Rosa (Consiglio Nazionale delle Ricerche Institute of Neuroscience, Milan, Italy), and F. Blasi (San Raffaele Institute, Milan, Italy), respectively.…”

Section: Methodsmentioning

confidence: 99%

The Rest Repression of the Neurosecretory Phenotype Is Negatively Modulated by BHC80, a Protein of the BRAF/HDAC Complex

Klajn¹,

Ferrai²,

Stucchi³

et al. 2009

J. Neurosci.

View full text Add to dashboard Cite

Expression of neurosecretion by nerve cells requires the levels of the transcription repressor element-1 silencing transcription factor (REST) to be very low. However, when high-REST clones of PC12 cells, defective of neurosecretion, were fused to other high-REST, non-neurosecretory cells, some neurosecretion was recovered. To clarify the mechanism of this recovery, we fused defective PC12 cells with human lymphocytes. A cytogenetic analysis revealed all hybrid clones that recovered neurosecretion to contain a fragment of chromosome 11 including the gene encoding BHC80, a protein of one of the complexes that mediate REST repression. In these clones, REST levels were as high as in defective PC12, whereas BHC80, localized in the nucleus, was 4-to 5-fold higher. Transient transfection of defective PC12 with various amounts of BHC80 cDNA induced (1) in defective PC12, the reexpression of only neurosecretion mRNAs; (2) in defective PC12 cotransfected with the REST negative construct DNA-binding domain (to attenuate gene repression), the recovery of a weak, but complete neurosecretory phenotype, including dense-core granules and their regulated exocytosis. Chromatin immunoprecipitation and immunodepletion analyses revealed the extensive BHC80 association with REST at the genes of two neurosecretion proteins, chromograninB and SNAP25, however only in the low-REST PC12, whereas in high-REST defective PC12 no association was appreciable. In defective PC12 transfected with BHC80 some association was reestablished. Therefore, the recovery of neurosecretion observed after fusion/transfection of defective PC12 depends on the reciprocal level of BHC80 and REST, with BHC80 working as a negative modulator of REST repression. This role appears of possible cell physiological and pathological importance.

show abstract

“…Moreover, we find additional subunits that contain plant homeodomain reminiscent of the BHC80 subunit in human complexes (17,21,30,33). Of notable exception is the absence of SWIRM-associated subunits containing the SANT domain.…”

Section: Discussionmentioning

confidence: 86%

Fission Yeast Homologs of Human Histone H3 Lysine 4 Demethylase Regulate a Common Set of Genes with Diverse Functions

Nicolas

Lee

Hakimi

et al. 2006

Journal of Biological Chemistry

View full text Add to dashboard Cite

Schizosaccharomyces pombe contains two proteins, SWIRM1 and SWIRM2, with close homology to human histone H3 lysine 4 demethylase. Both proteins contain the amino oxidase catalytic domain and a recently described DNA interaction SWIRM domain. Here we describe the biochemical isolation and the functional characterization of SWIRM1 and SWIRM2. Our results indicate that while SWIRM2 is an essential gene, cells lacking SWIRM1 are viable. We found that SWIRM1 and SWIRM2 are stably associated in a multiprotein complex, but intriguingly, unlike their human counterpart, S. pombe SWIRM complex contains neither a histone deacetylase nor any detectable demethylase activity. Genome-wide chromatin immunoprecipitation unexpectedly showed the absence of both SWIRM proteins from heterochromatic domains. Instead, consistent with biochemical analyses, SWIRM1 and SWIRM2 co-localize to a common set of target gene promoters whose functions are implicated in diverse processes including mitochondrial metabolism and transcriptional regulation. Importantly, we show that SWIRM1 is not only required for optimum transcription of its target genes but also display a global role in regulation of antisense transcription.Histones, the building blocks of chromatin in eukaryotic cells, are subject to several posttranslational modifications including methylation, acetylation, phosphorylation, and ubiquitylation (1). These histone modifications play a central role in modulating chromatin structure and function (2, 3). Of particular interest in recent years, histone methylation is known to impact several cellular processes such as chromatin organization, genomic imprinting, and transcriptional regulation (4 -6). This modification occurs on five lysine (K) residues of histone H3 and one lysine residue of histone H4 that can be methylated in three different modes, i.e. mono-, di-, or trimethylation. Generally, methylated H3K4, H3K36, and H3K79 residues are thought to be activation marks, whereas methylated H3K9, H3K27, and H4K20 residues represent repressive marks (4, 7). Arginine residues can also be methylated to generally lead to transcriptional activation (5).Although other histone modifications such as acetylation, phosphorylation, and ubiquitylation have been known to be reversible, histone methylation was considered irreversible. However, recent discoveries of several histone demethylases that can reverse methylated lysine and arginine residues have altered our views of histone demethylation (8 -13). PAD4/ PADI4 was the first reported histone arginine demethylase that demethyliminates monomethylarginine to produce citrulline (8, 9). BHC110/LSD1 has been identified as the first histone lysine demethylase that removes mono-or dimethyl-H3K4 by oxidation-based demethylation (13). More recently, a new family of histone demethylases was discovered to contain a JmjC domain capable of removing methyl groups on lysine residues by hydroxylation-based demethylation (10 -12).We and others (14 -20) have purified BHC110 as a component of several multiprotein ...

show abstract

Characterization of BHC80 in BRAF–HDAC complex, involved in neuron-specific gene repression

Cited by 51 publications

References 35 publications

Proteomics Analysis of Ring1B/Rnf2 Interactors Identifies a Novel Complex with the Fbxl10/Jhdm1B Histone Demethylase and the Bcl6 Interacting Corepressor

Proteomics Analysis of Ring1B/Rnf2 Interactors Identifies a Novel Complex with the Fbxl10/Jhdm1B Histone Demethylase and the Bcl6 Interacting Corepressor

The Rest Repression of the Neurosecretory Phenotype Is Negatively Modulated by BHC80, a Protein of the BRAF/HDAC Complex

Fission Yeast Homologs of Human Histone H3 Lysine 4 Demethylase Regulate a Common Set of Genes with Diverse Functions

Contact Info

Product

Resources

About