Assaying Chromatin Sirtuins

Zhong, Lei; Martínez-Pastor, Bárbara; Silberman, Dafne M.; Sebastián, Carlos; Mostoslavsky, Raúl

doi:10.1007/978-1-62703-637-5_10

Cited by 6 publications

(6 citation statements)

References 4 publications

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“…Chromatin acid extraction was performed as described previously ( Zhong et al, 2013 ). The mESCs or MEFs were collected and washed with PBS and resuspended with lysis buffer (10 mM HEPES [pH 7.4], 10 mM KCl, 0.05% NP-40, 1 mM sodium orthovanadate, and protease inhibitor cocktail [Roche]).…”

Section: Methodsmentioning

confidence: 99%

Inositol polyphosphate multikinase physically binds to the SWI/SNF complex and modulates BRG1 occupancy in mouse embryonic stem cells

Beon

Han

Yang

et al. 2022

eLife

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Inositol polyphosphate multikinase (IPMK), a key enzyme in inositol polyphosphate (IP) metabolism, is a pleiotropic signaling factor involved in major biological events, including transcriptional control. In the yeast, IPMK and its IP products promote the activity of the chromatin remodeling complex SWI/SNF, which plays a critical role in gene expression by regulating chromatin accessibility. However, the direct link between IPMK and chromatin remodelers remains unclear, raising the question of how IPMK contributes to transcriptional regulation in mammals. By employing unbiased screening approaches and in vivo/in vitro immunoprecipitation, here we demonstrate that mammalian IPMK physically interacts with the SWI/SNF complex by directly binding to SMARCB1, BRG1, and SMARCC1. Furthermore, we identified the specific domains required for IPMK-SMARCB1 binding. Notably, using CUT&RUN and ATAC-seq assays, we discovered that IPMK co-localizes with BRG1 and regulates BRG1 localization as well as BRG1-mediated chromatin accessibility in a genome-wide manner in mouse embryonic stem cells. Together, these findings show that IPMK regulates the promoter targeting of the SWI/SNF complex, thereby contributing to SWI/SNF-meditated chromatin accessibility, transcription, and differentiation in mouse embryonic stem cells.

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Section: Methodsmentioning

confidence: 99%

Inositol polyphosphate multikinase physically binds to the SWI/SNF complex and modulates BRG1 occupancy in mouse embryonic stem cells

Beon

Han

Yang

et al. 2022

eLife

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“…The protocol for the acid extraction of chromatin proteins was adapted from the Mostoslavsky lab protocol and performed as previously described 62,63 . Cell pellets were suspended with lysis buffer (10 mM HEPES pH 7.4, 10 mM KCl, 0.05% NP-40) three times the pellet volume and incubated on ice for 20 min.…”

Section: Methodsmentioning

confidence: 99%

CTDP1 regulates breast cancer survival and DNA repair through BRCT-specific interactions with FANCI

Krieger

Lagundžin

et al. 2019

Cell Death Discov.

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BRCA1 C-terminal domains are found in a specialized group of 23 proteins that function in the DNA damage response to protect genomic integrity. C-terminal domain phosphatase 1 (CTDP1) is the only phosphatase with a BRCA1 C-terminal domain in the human proteome, yet direct participation in the DNA damage response has not been reported. Examination of the CTDP1 BRCA1 C-terminal domain-specific protein interaction network revealed 103 high confidence interactions enriched in DNA damage response proteins, including FANCA and FANCI that are central to the Fanconi anemia DNA repair pathway necessary for the resolution of DNA interstrand crosslink damage. CTDP1 expression promotes DNA damage-induced FANCA and FANCD2 foci formation and enhances homologous recombination repair efficiency. CTDP1 was found to regulate multiple aspects of FANCI activity, including chromatin localization, interaction with γ-H2AX, and SQ motif phosphorylations. Knockdown of CTDP1 increases MCF-10A sensitivity to DNA interstrand crosslinks and double-strand breaks, but not ultraviolet radiation. In addition, CTDP1 knockdown impairs in vitro and in vivo growth of breast cancer cell lines. These results elucidate the molecular functions of CTDP1 in Fanconi anemia interstrand crosslink repair and identify this protein as a potential target for breast cancer therapy.

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“…Cell fractionation was performed according to previously published protocols with modifications (30,31). A fraction of the harvested cells was used to make whole cell extracts.…”

Section: Ada3mentioning

confidence: 99%

Alteration/Deficiency in Activation 3 (ADA3) Protein, a Cell Cycle Regulator, Associates with the Centromere through CENP-B and Regulates Chromosome Segregation

Mohibi¹,

Srivastava²,

Wang-France³

et al. 2015

Journal of Biological Chemistry

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Background: ADA3, a conserved component of several HAT complexes, regulates mitosis. Results: ADA3 associates with the centromere through CENP-B and regulates chromosome segregation by directing centromeric loading of CENP-B. Conclusion: ADA3 regulates mitosis by its association with the CENP-B/centromere and regulates segregation of chromosomes. Significance: This study provides the mechanism of how ADA3 regulates mitosis to maintain genomic stability.

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Assaying Chromatin Sirtuins

Cited by 6 publications

References 4 publications

Inositol polyphosphate multikinase physically binds to the SWI/SNF complex and modulates BRG1 occupancy in mouse embryonic stem cells

Inositol polyphosphate multikinase physically binds to the SWI/SNF complex and modulates BRG1 occupancy in mouse embryonic stem cells

CTDP1 regulates breast cancer survival and DNA repair through BRCT-specific interactions with FANCI

Alteration/Deficiency in Activation 3 (ADA3) Protein, a Cell Cycle Regulator, Associates with the Centromere through CENP-B and Regulates Chromosome Segregation

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