Cyclic AMP‐induced loss of histone H3 phosphorylation and disruption of cell cycle progression mediated through a novel cAMP binding protein

Abstract Background Forkhead transcription factors control cell growth in multiple cancer types. Foxd1 is essential for kidney development and mitochondrial metabolism, but its significance in renal cell carcinoma (ccRCC) has not been reported. Methods Transcriptome data from the TCGA database was used to correlate FOXD1 expression with patient survival. FOXD1 was knocked out in the 786-O cell line and known targets were analyzed. Reduced cell growth was observed and investigated in vitro using growth rate and Seahorse XF metabolic assays and in vivo using a xenograft model. Cell cycle characteristics were determined by flow cytometry and immunoblotting. Immunostaining for TUNEL and γH2AX was used to measure DNA damage. Association of the FOXD1 pathway with cell cycle progression was investigated through correlation analysis using the TCGA database. Results FOXD1 expression level in ccRCC correlated inversely with patient survival. Knockout of FOXD1 in 786-O cells altered expression of FOXD1 targets, particularly genes involved in metabolism (MICU1) and cell cycle progression. Investigation of metabolic state revealed significant alterations in mitochondrial metabolism and glycolysis, but no net change in energy production. In vitro growth rate assays showed a significant reduction in growth of 786-OFOXD1null. In vivo, xenografted 786-OFOXD1null showed reduced capacity for tumor formation and reduced tumor size. Cell cycle analysis showed that 786-OFOXD1null had an extended G2/M phase. Investigation of mitosis revealed a deficiency in phosphorylation of histone H3 in 786-OFOXD1null, and increased DNA damage. Genes correlate with FOXD1 in the TCGA dataset associate with several aspects of mitosis, including histone H3 phosphorylation. Conclusions We show that FOXD1 regulates the cell cycle in ccRCC cells by control of histone H3 phosphorylation, and that FOXD1 expression governs tumor formation and tumor growth. Transcriptome analysis supports this role for FOXD1 in ccRCC patient tumors and provides an explanation for the inverse correlation between tumor expression of FOXD1 and patient survival. Our findings reveal an important role for FOXD1 in maintaining chromatin stability and promoting cell cycle progression and provide a new tool with which to study the biology of FOXD1 in ccRCC.

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Histone H3 phosphorylation is required for the initiation, but not maintenance, of mammalian chromosome condensation

Journal of Cell Science ◽

10.1242/jcs.111.23.3497 ◽

1998 ◽

Vol 111 (23) ◽

pp. 3497-3506 ◽

Cited By ~ 12

Author(s):

A. Van Hooser ◽

D.W. Goodrich ◽

C.D. Allis ◽

B.R. Brinkley ◽

M.A. Mancini

Keyword(s):

Cell Cycle Progression ◽

Histone H3 ◽

Chromosome Morphology ◽

Chromosome Condensation ◽

S Phase ◽

Condensed State ◽

Cycle Progression ◽

H3 Phosphorylation ◽

Excess Substrate ◽

Histone H3 Phosphorylation

The temporal and spatial patterns of histone H3 phosphorylation implicate a specific role for this modification in mammalian chromosome condensation. Cells arrest in late G2 when H3 phosphorylation is competitively inhibited by microinjecting excess substrate at mid-S-phase, suggesting a requirement for activity of the kinase that phosphorylates H3 during the initiation of chromosome condensation and entry into mitosis. Basal levels of phosphorylated H3 increase primarily in late-replicating/early-condensing heterochromatin both during G2 and when premature chromosome condensation is induced. The prematurely condensed state induced by okadaic acid treatment during S-phase culminates with H3 phosphorylation throughout the chromatin, but in an absence of mitotic chromosome morphology, indicating that the phosphorylation of H3 is not sufficient for complete condensation. Mild hypotonic treatment of cells arrested in mitosis results in the dephosphorylation of H3 without a cytological loss of chromosome compaction. Hypotonic-treated cells, however, complete mitosis only when H3 is phosphorylated. These observations suggest that H3 phosphorylation is required for cell cycle progression and specifically for the changes in chromatin structure incurred during chromosome condensation.

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P2-438: Amyloid precursor protein-binding protein 1(APP-BP1) is critically required for the cell cycle progression in neural stem cells

Alzheimer s & Dementia ◽

10.1016/j.jalz.2008.05.1517 ◽

2008 ◽

Vol 4 ◽

pp. T502-T502

Author(s):

Yuyoung Joo ◽

Sung Ji Ha ◽

Sang Hyung Lee ◽

Yoo-Hun Suh ◽

Hye-Sun Kim

Keyword(s):

Cell Cycle ◽

Stem Cells ◽

Neural Stem Cells ◽

Amyloid Precursor Protein ◽

Protein Binding ◽

Cell Cycle Progression ◽

Binding Protein ◽

Precursor Protein ◽

Cycle Progression

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The GTP-binding Protein Rho1p Is Required for Cell Cycle Progression and Polarization of the Yeast Cell

The Journal of Cell Biology ◽

10.1083/jcb.146.2.373 ◽

1999 ◽

Vol 146 (2) ◽

pp. 373-387 ◽

Cited By ~ 39

Author(s):

Jana Drgonová ◽

Tomás Drgon ◽

Dong-Hyun Roh ◽

Enrico Cabib

Keyword(s):

Cell Cycle ◽

Yeast Cell ◽

Cell Cycle Progression ◽

Binding Protein ◽

Cell Polarization ◽

Nonpermissive Temperature ◽

Gtp Binding Protein ◽

Cycle Progression ◽

Glucan Synthase ◽

Gtp Binding

Previous work showed that the GTP-binding protein Rho1p is required in the yeast, Saccharomyces cerevisiae, for activation of protein kinase C (Pkc1p) and for activity and regulation of β(1→3)glucan synthase. Here we demonstrate a hitherto unknown function of Rho1p required for cell cycle progression and cell polarization. Cells of mutant rho1E45I in the G1 stage of the cell cycle did not bud at 37°C. In those cells actin reorganization and recruitment to the presumptive budding site did not take place at the nonpermissive temperature. Two mutants in adjacent amino acids, rho1V43T and rho1F44Y, showed a similar behavior, although some budding and actin polarization occurred at the nonpermissive temperature. This was also the case for rho1E45I when placed in a different genetic background. Cdc42p and Spa2p, two proteins that normally also move to the bud site in a process independent from actin organization, failed to localize properly in rho1E45I. Nuclear division did not occur in the mutant at 37°C, although replication of DNA proceeded slowly. The rho1 mutants were also defective in the formation of mating projections and in congregation of actin at the projections in the presence of mating pheromone. The in vitro activity of β(1→3)glucan synthase in rho1 E45I, although diminished at 37°C, appeared sufficient for normal in vivo function and the budding defect was not suppressed by expression of a constitutively active allele of PKC1. Reciprocally, when Pkc1p function was eliminated by the use of a temperature-sensitive mutation and β(1→3)glucan synthesis abolished by an echinocandin-like inhibitor, a strain carrying a wild-type RHO1 allele was able to produce incipient buds. Taken together, these results reveal a novel function of Rho1p that must be executed in order for the yeast cell to polarize.

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The TIA1 RNA-Binding Protein Family Regulates EIF2AK2-Mediated Stress Response and Cell Cycle Progression

Molecular Cell ◽

10.1016/j.molcel.2018.01.011 ◽

2018 ◽

Vol 69 (4) ◽

pp. 622-635.e6 ◽

Cited By ~ 28

Author(s):

Cindy Meyer ◽

Aitor Garzia ◽

Michael Mazzola ◽

Stefanie Gerstberger ◽

Henrik Molina ◽

...

Keyword(s):

Cell Cycle ◽

Stress Response ◽

Cell Cycle Progression ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Protein Family ◽

Cycle Progression

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D-site binding protein regulates cell proliferation through mediating cell cycle progression in rat mesangial cells

Tissue and Cell ◽

10.1016/j.tice.2019.08.006 ◽

2019 ◽

Vol 61 ◽

pp. 35-43

Author(s):

Hongli Jiang ◽

Jie Li ◽

Xin He ◽

Jinhong Xue ◽

Shanshan Liang ◽

...

Keyword(s):

Cell Cycle ◽

Cell Proliferation ◽

Cell Cycle Progression ◽

Mesangial Cells ◽

Binding Protein ◽

Cycle Progression ◽

Site Binding

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Characterization of HDACI OSU42 as a Novel Histone Deacetylase Inhibitor in AML Cell Lines.

Blood ◽

10.1182/blood.v108.11.1988.1988 ◽

2006 ◽

Vol 108 (11) ◽

pp. 1988-1988

Author(s):

Jin Sun ◽

Shujun Liu ◽

Jianhua Yu ◽

Min Wei ◽

Charlene Mao ◽

...

Keyword(s):

Cell Cycle ◽

Gene Silencing ◽

Cell Cycle Progression ◽

Chromatin Condensation ◽

Regulation Of Gene Expression ◽

Histone H3 ◽

Malignant Cells ◽

Cycle Progression ◽

Nb4 Cells ◽

Histone Hypoacetylation

Abstract Histone acetylation plays a key role in the regulation of gene expression. Histone hyperacetylation is associated with chromatin opening and gene transcription, while histone hypoacetylation is associated with chromatin condensation and gene silencing. Abnormal histone hypoacetylation mediated by aberrant activity of histone deacetylases (HDACs) has been found to be associated with silencing of tumor suppressor and growth inhibitory genes in malignant cells. HDAC inhibitors (HDACIs) can relieve HDAC-mediated gene silencing and thereby induce normal patterns of cell cycle, differentiation and apoptosis in malignant cells. HDACI OSU 42 is a novel hydroxamate tethered phenylbutyrate derivative that was designed and synthesized at our institution, and exhibited IC50s at submicromolar level, compared with millimolar level for other members of this classes of HDACIs such as valproic acid (VPA). We characterized the activity of this compound in acute myeloid leukemia (AML) cells. It is known that the fusion proteins AML1/ETO and PML / RAR alpha that characterized t(8;21) and t(15;17) AML silence target genes through recruitment of HDACs to their promoter regions. Therefore we utilized AML1/ETO-positive Kasumi-1 and PML/RARA-positive NB4 cells to test the activity of HDACI OSU 42 and used THP-1 cells, characterized by AF9/MLL fusion gene, as a control. We hypothesized that by virtue of the fusion genes, Kasumi-1 and NB4 are more susceptible to HDACI treatment. IC50s for proliferation inhibition in Kasumi-1 cells treated with HDACI OSU42 were 71.8±14.3nM for 24hr and 31.3± 0.4nM for 48hr, significantly lower than VPA (2.0mM for 24hr, 0.9mM for 48hr). The IC50s for NB4 were 237.7±6.5nM for 24hr and 119±6.4nM for 48hr. As a contrast, IC50 for THP-1 was 507.3±68.3nM for 48hr. HDACI OSU42 inhibited 80% of total HDAC activity at 125nM in both Kasumi-1 and NB4; 30nM HDACI OSU42 induced hyperacetylation of histone H3 and H4. Apoptosis analysis showed that nearly 60% more of Kasumi-1 and NB4 underwent apoptosis after treated with 1μM of HDACI OSU42 for 24hr, compared with their untreated control. On the other hand, the same treatment only induced 15% more of THP-1 undergoing apoptosis. Apoptotic effect of HDACI OSU42 was mediated by activation of caspase 9 and caspase 3. Cell cycle analysis demonstrated that treatment of Kasumi-1 and NB4 with 150nM of HDACI OSU 42 inhibited cell cycle progression and arrested 20% to 30% more cells at S phase or G2/M phase, whereas this treatment had not effect on cell cycle progression of THP-1. This was consistent with the up-regulated expression of p21 at both transcription level and protein level. Q-PCR data suggested that Kasumi-1 and NB4 treated with HDACI OSU42 expressed ~10 folds of p21 higher than untreated cells. Chromatin immunoprecipitation assay revealed 10 to 50 folds increase in acetylation level of histone H3 and H4 associated with p21 promoter. Kasumi-1 and NB4 cells also show differentiation ability (increase in CD14 and CD 13 expression by flow cytometry) when treated with 30nM of HDACI OSU42, whereas THP-1 remained undifferentiated. These results support the activity of HDACI OSU42 as a new potent HDACI in AML.

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The possible role of cyclic AMP in the growth and cell cycle progression in the dinoflagellate Crypthecodinium cohnii (Biecheler)

10.14711/thesis-b647128 ◽

1999 ◽

Author(s):

Mo Ching Lam

Keyword(s):

Cell Cycle ◽

Cyclic Amp ◽

Cell Cycle Progression ◽

Cycle Progression ◽

Crypthecodinium Cohnii

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Cell Cycle-dependent Expression and Nucleolar Localization of hCAP-H

Molecular Biology of the Cell ◽

10.1091/mbc.12.11.3527 ◽

2001 ◽

Vol 12 (11) ◽

pp. 3527-3537 ◽

Cited By ~ 32

Author(s):

Olga A. Cabello ◽

Elena Eliseeva ◽

WeiGong He ◽

Hagop Youssoufian ◽

Sharon E. Plon ◽

...

Keyword(s):

Cell Cycle ◽

Histone H3 ◽

Chromosome Condensation ◽

Nucleolar Localization ◽

Mitotic Chromosomes ◽

Proliferating Cells ◽

Protein Levels ◽

Sister Chromatids ◽

H3 Phosphorylation ◽

Histone H3 Phosphorylation

Condensin is a conserved 13S heteropentamer composed of two nonidentical structural maintenance of chromosome (SMC) family proteins, in Xenopus XCAP-C and XCAP-E, and three regulatory subunits, XCAP-D2, XCAP-G, and XCAP-H. Both biochemical and genetic analyses have demonstrated an essential role for the 13S condensin complex in mitotic chromosome condensation. Further, a potential requirement for condensin in completion of chromatid arm separation in early anaphase is demonstrated by the mutational phenotypes of the Drosophila homologues ofXCAP-H, barren and XCAP-C,DmSMC4. In this study we have investigated the expression and subcellular distribution of hCAP-H, the human homolog of XCAP-H, in order to better understand its cellular functions. Transcription of hCAP-H was restricted to proliferating cells with highest expression during the G2 phase of the cell cycle. In contrast, cellular hCAP-H protein levels were constant throughout the cell cycle. hCAP-H was found to be associated with mitotic chromosomes exhibiting a nonuniform but symmetric distribution along sister chromatids. The symmetry of hCAP-H association with sister chromatids suggests that there are sequence-dependent domains of condensin aggregation. During interphase hCAP-H, -C, and -E, have distinct punctate nucleolar localization, suggesting that condensin may associate with and modulate the conformation and function of rDNA. hCAP-H association with condensed chromatin was not observed in the early phase of chromosome condensation when histone H3 phosphorylation has already taken place. This finding is consistent with the hypothesis that histone H3 phosphorylation precedes condensin-mediated condensation.

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