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

Mohibi, Shakur; Srivastava, Shashank; Wang-France, Jun; Mirza, Sameer; Zhao, Xiangshan; Band, Hamid; Band, Vimla

doi:10.1074/jbc.m115.685511

Cited by 12 publications

(12 citation statements)

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“…For example, we identified ADA3 as a novel human papillomavirus E6 oncoprotein-binding protein (17), and additional studies revealed that ADA3 binds to nuclear hormone receptors, such as estrogen receptor and retinoid acid receptor, and enhances their transcriptional activation function (8,(18)(19)(20)(21). Recent studies have identified an essential role of ADA3 in normal cell cycle progression and maintenance of genomic stability (5,13,22,23).…”

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“…GST pulldown assay was performed as described previously (23). Briefly, 1 g of bacterially purified GST or GST-ADA3 protein bound to beads was used as bait and incubated with 300 ng of baculovirally purified SIRT1 protein, purchased from R&D Systems (catalog no.…”

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confidence: 99%

“…Rescue proliferation assays for various acetylation-defective mutants were performed as previously described (23). Briefly, Ada3 FL/FL MEFs stably overexpressing empty vector, ADA3 WT or the K418R, 7KR, or 8KR mutant were plated in triplicates in 96-well plates (150 cells/well) 24 h after infection with adenovirus EGFP or EGFP-Cre.…”

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Acetylation of Mammalian ADA3 Is Required for Its Functional Roles in Histone Acetylation and Cell Proliferation

Mohibi

Srivastava

Bele

et al. 2016

Molecular and Cellular Biology

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Alteration/deficiency in activation 3 (ADA3) is an essential component of specific histone acetyltransferase (HAT) complexes. We have previously shown that ADA3 is required for establishing global histone acetylation patterns and for normal cell cycle progression (S. Mohibi et al., J Biol Chem 287:29442-29456, 2012, http://dx.doi.org/10.1074/jbc.M112.378901). Here, we report that these functional roles of ADA3 require its acetylation. We show that ADA3 acetylation, which is dynamically regulated in a cell cycle-dependent manner, reflects a balance of coordinated actions of its associated HATs, GCN5, PCAF, and p300, and a new partner that we define, the deacetylase SIRT1. We use mass spectrometry and site-directed mutagenesis to identify major sites of ADA3 acetylated by GCN5 and p300. Acetylation-defective mutants are capable of interacting with HATs and other components of HAT complexes but are deficient in their ability to restore ADA3-dependent global or locus-specific histone acetylation marks and cell proliferation in Ada3-deleted murine embryonic fibroblasts (MEFs). Given the key importance of ADA3-containing HAT complexes in the regulation of various biological processes, including the cell cycle, our study presents a novel mechanism to regulate the function of these complexes through dynamic ADA3 acetylation.A lteration/deficiency in activation 3 (ADA3) protein is a conserved component of key chromatin-modifying complexes that contain either GCN5 or PCAF histone acetyl transferases (HATs), such as SAGA (Spt/Ada/Gcn5) in yeast, ATAC (ADA2a-containing complex), STAGA (SPT3/TAFII31/GCN5 acetyltransferase), and TFTC (TATA binding protein free-TAF containing complex) in metazoans (1-7). Within these complexes, ADA3 associates with GCN5 and ADA2 to form the HAT module. ADA3 has also been shown to associate with p300, the most well-defined HAT of mammalian systems (8, 9). ADA3 is essential for the HAT activity of p300-and GCN5-containing HAT complexes toward histones (10-14) as well as of nonhistone proteins, such as p53 and ␤-catenin (15, 16).Although strongly implicated in the regulation of HAT activity of ADA3-containing complexes, additional functions for ADA3 have been reported. For example, we identified ADA3 as a novel human papillomavirus E6 oncoprotein-binding protein (17), and additional studies revealed that ADA3 binds to nuclear hormone receptors, such as estrogen receptor and retinoid acid receptor, and enhances their transcriptional activation function (8,(18)(19)(20)(21). Recent studies have identified an essential role of ADA3 in normal cell cycle progression and maintenance of genomic stability (5, 13, 22, 23).Whether ADA3's role in these processes is merely a passive structural one or is actively regulated is unknown. Posttranslational modification represents one potential mechanism to regulate ADA3 function, and in fact yeast ADA3 was found to be modified by acetylation (24). Consistent with this idea, we observed that human ADA3 is also acetylated in vitro by its interacting HAT p300 (13). H...

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Acetylation of Mammalian ADA3 Is Required for Its Functional Roles in Histone Acetylation and Cell Proliferation

Mohibi

Srivastava

Bele

et al. 2016

Molecular and Cellular Biology

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“…Virally transduced cells were selected in 0.5 μg/ml puromycin for 3 days, and expression of endogenous ADA3 was assessed in the whole cell lysate using Western blotting with anti-ADA3 antibody. As an additional approach ADA3 was depleted by using siRNA (sc-7846) and control siRNA (sc-37007) purchased from Santa Cruz Biotechnology, using the same protocol as described in reference [5]. …”

Section: Methodsmentioning

confidence: 99%

“…Thus, a better understanding of cell cycle machinery and its aberrations in cancer are of fundamental importance in cell and cancer biology. Recently ADA3, a component of histone acetyltransferase (HAT) complexes, has emerged as a key regulator of cell cycle progression through G1/S and G2/M transitions and in maintaining the genomic stability by governing the faithful segregation of chromosomes [3–5]. …”

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ADA3 regulates normal and tumor mammary epithelial cell proliferation through c-MYC

et al. 2016

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BackgroundWe have established the critical role of ADA3 as a coactivator of estrogen receptor (ER), as well as its role in cell cycle progression. Furthermore, we showed that ADA3 is predominantly nuclear in mammary epithelium, and in ER+, but is cytoplasmic in ER- breast cancers, the latter correlating with poor survival. However, the role of nuclear ADA3 in human mammary epithelial cells (hMECs), and in ER+ breast cancer cells, as well as the importance of ADA3 expression in relation to patient prognosis and survival in ER+ breast cancer have remained uncharacterized.MethodsWe overexpressed ADA3 in hMECs or in ER+ breast cancer cells and assessed the effect on cell proliferation. The expression of ADA3 was analyzed then correlated with the expression of various prognostic markers, as well as survival of breast cancer patients.ResultsOverexpression of ADA3 in ER- hMECs as well as in ER+ breast cancer cell lines enhanced cell proliferation. These cells showed increased cyclin B and c-MYC, decreased p27 and increased SKP2 levels. This was accompanied by increased mRNA levels of early response genes c-FOS, EGR1, and c-MYC. Analysis of breast cancer tissue specimens showed a significant correlation of ADA3 nuclear expression with c-MYC expression. Furthermore, nuclear ADA3 and c-MYC expression together showed significant correlation with tumor grade, mitosis, pleomorphism, NPI, ER/PR status, Ki67 and p27 expression. Importantly, within ER+ cases, expression of nuclear ADA3 and c-MYC also significantly correlated with Ki67 and p27 expression. Univariate Kaplan Meier analysis of four groups in the whole, as well as the ER+ patients showed that c-MYC and ADA3 combinatorial phenotypes showed significantly different breast cancer specific survival with c-MYC-high and ADA3-Low subgroup had the worst outcome. Using multivariate analyses within the whole cohort and the ER+ subgroups, the significant association of ADA3 and c-MYC expression with patients’ outcome was independent of tumor grade, stage and size, and ER status.ConclusionADA3 overexpression enhances cell proliferation that is associated with increased expression of c-MYC. Expression patterns with respect to ADA3/c-MYC can divide patients into four significantly different subgroups, with c-MYC High and ADA3 Low status independently predicting poor survival in patients.Electronic supplementary materialThe online version of this article (doi:10.1186/s13058-016-0770-9) contains supplementary material, which is available to authorized users.

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DNA Sequences in Centromere Formation and Function

Dumont

Fachinetti

2017

Progress in Molecular and Subcellular Biology

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Faithful chromosome segregation during cell division depends on the centromere, a complex DNA/protein structure that links chromosomes to spindle microtubules. This chromosomal domain has to be marked throughout cell division and its chromosomal localization preserved across cell generations. From fission yeast to human, centromeres are established on a series of repetitive DNA sequences and on specialized centromeric chromatin. This chromatin is enriched with the histone H3 variant, named CENP-A, that was demonstrated to be the epigenetic mark that maintains centromere identity and function indefinitely. Although centromere identity is thought to be exclusively epigenetic, the presence of specific DNA sequences in the majority of eukaryotes and of the centromeric protein CENP-B that binds to these sequences, suggests the existence of a genetic component as well. In this review, we will highlight the importance of centromeric sequences for centromere formation and function, and discuss the centromere DNA sequence/CENP-B paradox.

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

Cited by 12 publications

References 53 publications

Acetylation of Mammalian ADA3 Is Required for Its Functional Roles in Histone Acetylation and Cell Proliferation

Acetylation of Mammalian ADA3 Is Required for Its Functional Roles in Histone Acetylation and Cell Proliferation

ADA3 regulates normal and tumor mammary epithelial cell proliferation through c-MYC

DNA Sequences in Centromere Formation and Function

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