PIAS1 binds p300 and behaves as a coactivator or corepressor of the transcription factor c-Myb dependent on SUMO-status

Ledsaak, Marit; Bengtsen, Mads; Molværsmyr, Ann-Kristin; Fuglerud, Bettina M.; Matre, Vilborg; Eskeland, Ragnhild; Gabrielsen, Odd S.

doi:10.1016/j.bbagrm.2016.03.011

Cited by 14 publications

(19 citation statements)

References 77 publications

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“…The cells were cultured as described in ( 32 ) and K562 cells stably expressing TY-tagged c-Myb variants (empty vector, 3xTY1-c-Myb, 3xTY-c-Myb D152V) were generated as described in ( 33 ). For transfection of K562 cells we used Ingenio Electroporation Solution (Mirus Bio) following the manufacture's guidelines.…”

Section: Methodsmentioning

confidence: 99%

“…Twenty-four hours after transfection COS-1 cells were lysed in interaction buffer (20 mM HEPES pH 7.6, 10% glycerol, 0.2% Triton-X-100, 150 mM KAc, 1 mM Dithiothreitol (DTT)) supplemented with Complete protease inhibitor cocktail (Roche) and used in GST pulldown assays. Luciferase assays were performed as described in ( 33 ) using a Wallac Victor2 multi-plate reader (Perkin Elmer). Plasmids used are described in Supplementary Materials and Methods .…”

Section: Methodsmentioning

confidence: 99%

“…GST and GST fusion proteins were expressed in E. coli as previously described ( 54 ). GST pulldown was performed as described in ( 33 ), but after pulldown the beads were washed twice in interaction buffer with 0.5% Triton-X-100. Proteins were detected by Western blotting as described in Supplementary Materials and Methods .…”

Section: Methodsmentioning

confidence: 99%

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A c-Myb mutant causes deregulated differentiation due to impaired histone binding and abrogated pioneer factor function

Fuglerud

Lemma

Wanichawan

et al. 2017

Nucleic Acids Research

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The transcription factor c-Myb is involved in early differentiation and proliferation of haematopoietic cells, where it operates as a regulator of self-renewal and multi-lineage differentiation. Deregulated c-Myb plays critical roles in leukaemias and other human cancers. Due to its role as a master regulator, we hypothesized it might function as a pioneer transcription factor. Our approach to test this was to analyse a mutant of c-Myb, D152V, previously reported to cause haematopoietic defects in mice by an unknown mechanism. Our transcriptome data from K562 cells indicates that this mutation specifically affects c-Myb's ability to regulate genes involved in differentiation, causing failure in c-Myb's ability to block differentiation. Furthermore, we see a major effect of this mutation in assays where chromatin opening is involved. We show that each repeat in the minimal DNA-binding domain of c-Myb binds to histones and that D152V disrupts histone binding of the third repeat. ATAC-seq data indicates this mutation impairs the ability of c-Myb to cause chromatin opening at specific sites. Taken together, our findings support that c-Myb acts as a pioneer factor and show that D152V impairs this function. The D152V mutant is the first mutant of a transcription factor specifically destroying pioneer factor function.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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A c-Myb mutant causes deregulated differentiation due to impaired histone binding and abrogated pioneer factor function

Fuglerud

Lemma

Wanichawan

et al. 2017

Nucleic Acids Research

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“…A HEK293 reporter cell line harboring a 5ϫGRE Gal4-luciferase reporter integrated transgene (26) was co-transfected with an HA-tagged Gal-fusion derivative of CHD3 and a 3ϫFLAG-tagged SENP1. The ChIP analysis was performed as described previously (61).…”

Section: Chipmentioning

confidence: 99%

The SUMO protease SENP1 and the chromatin remodeler CHD3 interact and jointly affect chromatin accessibility and gene expression

Rodríguez-Castañeda

Lemma

Cuervo

et al. 2018

Journal of Biological Chemistry

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The small ubiquitin-like modifier (SUMO) post-translationally modifies lysine residues of transcription factors and co-regulators and thereby contributes to an important layer of control of the activities of these transcriptional regulators. Likewise, deSUMOylation of these factors by the sentrin-specific proteases (SENPs) also plays a role in gene regulation, but whether SENPs functionally interact with other regulatory factors that control gene expression is unclear. In the present work, we focused on SENP1, specifically, on its role in activation of gene expression investigated through analysis of the SENP1 interactome, which revealed that SENP1 physically interacts with the chromatin remodeler chromodomain helicase DNA-binding protein 3 (CHD3). Using several additional methods, including GST pulldown and co-immunoprecipitation assays, we validated and mapped this interaction, and using CRISPR-Cas9-generated CHD3- and SENP1-KO cells (in the haploid HAP1 cell line), we investigated whether these two proteins are functionally linked in regulating chromatin remodeling and gene expression. Genome-wide ATAC-Seq analysis of the CHD3- and SENP1-KO cells revealed a large degree of overlap in differential chromatin openness between these two mutant cell lines. Moreover, motif analysis and comparison with ChIP-Seq profiles in K562 cells pointed to an association of CHD3 and SENP1 with CCCTC-binding factor (CTCF) and SUMOylated chromatin-associated factors. Lastly, genome-wide RNA-Seq also indicated that these two proteins co-regulate the expression of several genes. We propose that the functional link between chromatin remodeling by CHD3 and deSUMOylation by SENP1 uncovered here provides another level of control of gene expression.

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“…Sin embargo, el papel de SUMO como regulador transcripcional continúa siendo controvertido. Existen trabajos anteriores donde se ha demostrado que la SUMOilación de diferentes factores transcripcionales puede favorecer la represión transcripcional de sus genes diana [180][181][182] , así como otros donde se describe que promueve el reclutamiento de co-activadores transcripcionales como CBP y p300 [183][184][185] . Así pues, con la finalidad de evitar el uso de inhibidores químicos, se identificaron los residuos de lisina claves en el proceso de SUMOilación de N1ICD que se mutaron y sobre los que se realizaron estudios adicionales.…”

Section: Modulación De La Vía De Notch En Mscs En Presencia O Ausenciunclassified

Interacción entre las vías HIF, Notch y SUMO y su implicación en la biología de las hMSCs

Calduch¹

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PIAS1 binds p300 and behaves as a coactivator or corepressor of the transcription factor c-Myb dependent on SUMO-status

Cited by 14 publications

References 77 publications

A c-Myb mutant causes deregulated differentiation due to impaired histone binding and abrogated pioneer factor function

A c-Myb mutant causes deregulated differentiation due to impaired histone binding and abrogated pioneer factor function

The SUMO protease SENP1 and the chromatin remodeler CHD3 interact and jointly affect chromatin accessibility and gene expression

Interacción entre las vías HIF, Notch y SUMO y su implicación en la biología de las hMSCs

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