Nuclear c-Abl-mediated tyrosine phosphorylation induces chromatin structural changes through histone modifications that include H4K16 hypoacetylation

Aoyama, Kazumasa; Fukumoto, Yasunori; Ishibashi, Keiichiro; Kubota, Sho; Morinaga, Takao; Horiike, Yasuyoshi; Yuki, Ryuzaburo; Takahashi, Akinori; Nakayama, Yuji; Yamaguchi, Naoto

doi:10.1016/j.yexcr.2011.09.013

Cited by 30 publications

(69 citation statements)

References 73 publications

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“…19) In brief, cells were fixed in 4% paraformaldehyde for 20 min at room temperature and permeabilized in phosphate-buffered saline (PBS) containing 0.2% Triton X-100 and 3% bovine serum albumin. Cells were subsequently reacted with anti-Slug antibody (#9585) for 1 h, washed with PBS containing 0.1% saponin, stained with Alexa Fluor 488-conjugated secondary antibody for 1 h, and mounted with ProLong antifade reagent (Life Technologies).…”

Section: Methodsmentioning

confidence: 99%

FOXA1 Induces E-Cadherin Expression at the Protein Level via Suppression of Slug in Epithelial Breast Cancer Cells

Anzai

Hirata

Shibazaki

et al. 2017

Biological & Pharmaceutical Bulletin

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Epithelial-to-mesenchymal transition (EMT) is an important process during embryonic development and tumor progression by which adherent epithelial cells acquire mesenchymal properties. Forkhead box protein A1 (FOXA1) is a transcriptional regulator preferentially expressed in epithelial breast cancer cells, and its expression is lost in mesenchymal breast cancer cells. However, the implication of this biased expression of FOXA1 in breast cancer is not fully understood. In this study, we analyzed the involvement of FOXA1 in EMT progression in breast cancer, and found that stable expression of FOXA1 in the mesenchymal breast cancer MDA-MB-231 cells strongly induced the epithelial marker E-cadherin at the mRNA and protein levels. Furthermore, stable expression of FOXA1 was found to reduce the mRNA and protein expression of Slug, a repressor of E-cadherin expression. FOXA1 knockdown in the epithelial breast cancer MCF7 cells reduced E-cadherin protein expression without decreasing its mRNA expression. In addition, FOXA1 knockdown in MCF7 cells up-regulated Slug mRNA and protein expression. Notably, similar to FOXA1 knockdown, stable expression of Slug in MCF7 cells reduced E-cadherin protein expression without decreasing its mRNA expression. Taken together, these results suggest that although FOXA1 can induce E-cadherin mRNA expression, it preferentially promotes E-cadherin expression at the protein level by suppressing Slug expression in epithelial breast cancer, and that the balance of this FOXA1-Slug axis regulates EMT progression.Key words epithelial-mesenchymal transition; E-cadherin; forkhead box protein A1; Slug; estrogen receptor Epithelial-to-mesenchymal transition (EMT) is a process that converts adherent epithelial cells to motile mesenchymal cells during embryonic development. 1) EMT is also involved in tumor progression through inducing metastasis and chemoresistance in tumor cells.2-4) Therefore, a better understanding of the molecular mechanisms underlying EMT in tumor cells is beneficial for development of novel antitumor agents.E-Cadherin is a crucial protein that mediates cell-cell adhesion in epithelial cells.1) E-Cadherin expression is dramatically reduced during EMT, which causes loss of cell-cell adhesion and gain of cell motility. Reduction in E-cadherin expression is mainly caused by the transcriptional repressors of E-cadherin gene (CDH1).2,5) A major transcriptional repressor of CDH1 is the zinc finger factor Slug. Slug binds to the E-box sequences present in the enhancer region of CDH1 and represses its expression. 6)Forkhead box A1 (FOXA1) is a member of the forkhead box family of transcription factors. FOXA1 contains the forkhead DNA-binding domain and the N-and C-terminal transactivation domains.7) FOXA1 regulates expression of many genes involved in the development of various tissues, including mammary gland, liver, midbrain, and lung. 8) FOXA1 is also known to function as a chromatin remodeling factor that opens closed chromatin and facilitates the recruitment of other transcription fa...

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Section: Methodsmentioning

confidence: 99%

FOXA1 Induces E-Cadherin Expression at the Protein Level via Suppression of Slug in Epithelial Breast Cancer Cells

Anzai

Hirata

Shibazaki

et al. 2017

Biological & Pharmaceutical Bulletin

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“…The mechanism of Src-mediated suppression of apoptosis through Src substrates still remains poorly understood, possibly because protein-tyrosine phosphorylation is extremely unstable within cells. We, therefore, assume that phosphorylation levels of substrates are very rapidly regulated by a balance of the activities between tyrosine kinases and tyrosine phosphatases, like ON/OFF switching in a microprocessor (5,(22)(23)(24)(25)(26)(27)57). Nonetheless, carefully using a high dose of the potent tyrosine phosphatase inhibitor Na 3 VO 4 , we are able to detect tyrosine phosphorylation of endogenous proteins and have shown the significance of tyrosine-phosphorylated substrates (5,24,26,27).…”

Section: Discussionmentioning

confidence: 99%

“…Immunofluorescence-Confocal images were obtained using a Fluoview Fv500 confocal laser scanning microscope with a 20 ϫ 1.00 NA or 40 ϫ 1.00 NA objective (Olympus, Tokyo), as described (23,80). One planar (xy) section slice images (0.6-or 2.0-m thickness) were shown.…”

Section: Methodsmentioning

confidence: 99%

“…We have shown that Lyn, a Src member, is imported into and rapidly exported from the nucleus (21) and that nuclear tyrosine phosphorylation has a role in global changes of chromatin structure and histone modifications (13,(22)(23)(24). To explore in detail the roles of protein-tyrosine phosphorylation in cell functions, we recently performed phosphoproteomic analyses (5,25) and revealed novel roles for tyrosine phosphorylation of nuclear proteins, such as the heterochromatin protein KAP1 (KRAB-associated protein 1)/TIF1␤/TRIM28 (5), the transcription factors JunB (26) and FoxA1 (27), and the chromatinassociated protein AKAP8 (A-kinase anchoring protein 8) (24).…”

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Src Acts as an Effector for Ku70-dependent Suppression of Apoptosis through Phosphorylation of Ku70 at Tyr-530

Morii

Kubota

Honda

et al. 2017

Journal of Biological Chemistry

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Edited by Xiao-Fan WangSrc-family tyrosine kinases are widely expressed in many cell types and participate in a variety of signal transduction pathways. Despite the significance of Src in suppression of apoptosis, its mechanism remains poorly understood. Here we show that Src acts as an effector for Ku70-dependent suppression of apoptosis. Inhibition of endogenous Src activity promotes UV-induced apoptosis, which is impaired by Ku70 knockdown. Src phosphorylates Ku70 at Tyr-530, being close to the possible acetylation sites involved in promotion of apoptosis. Src-mediated phosphorylation of Ku70 at Tyr-530 decreases acetylation of Ku70, whereas Src inhibition augments acetylation of Ku70. Importantly, knockdown-rescue experiments with stable Ku70 knockdown cells show that the nonphosphorylatable Y530F mutant of Ku70 reduces the ability of Ku70 to suppress apoptosis accompanied by augmentation of Ku70 acetylation. Our results reveal that Src plays a protective role against hyperactive apoptotic cell death by reducing apoptotic susceptibility through phosphorylation of Ku70 at Tyr-530.

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“…16,20,21) Immunofluorescence Confocal images were obtained with a Fluoview FV500 confocal laser scanning microscope (Olympus, Tokyo, Japan), as described. 1) In brief, cells were fixed in 4% paraformaldehyde for 20 min at room temperature and permeabilized in phosphate-buffered saline (PBS) containing 0.1% saponin and 3% bovine serum albumin at room temperature. Cells were subsequently incubated with antibodies for 1 h, washed with PBS containing 0.1% saponin, stained with Alexa Fluor 488-or 546-conjugated secondary antibody for 1 h, and mounted with ProLong antifade reagent (Thermo Fisher Scientific).…”

Section: Highlighted Paper Selected By Editor-in-chiefmentioning

confidence: 99%

The Truncated Isoform of the Receptor Tyrosine Kinase ALK Generated by Alternative Transcription Initiation (ALKATI) Induces Chromatin Structural Changes in the Nucleus in a Kinase Activity-Dependent Manner

Takakura

Yamaguchi

Honda

et al. 2017

Biological & Pharmaceutical Bulletin

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Anaplastic lymphoma kinase (ALK) is a receptor-type tyrosine kinase that promotes cell growth upon stimulation with ligands such as midkine and pleiotrophin. Recently, a truncated isoform of ALK was identified in a variety of tumors. This isoform is expressed from a novel ALK transcript initiated from a de novo alternative transcription initiation (ATI) site in ALK intron 19 (referred to as ALK ATI ). ALK ATI , which consists of only the intracellular kinase domain, localizes to the nucleus as well as the cytoplasm. However, its nuclear role is unknown. In this study, we determined that ALK ATI promoted chromatin structural changes in the nucleus in a kinase activity-dependent manner. We found that expression of ALK ATI increased the level of the heterochromatin marker Lys9 tri-methylated histone H3. In addition, we demonstrated that ALK ATI phosphorylated the nuclear protein A-kinase anchoring protein 8 (AKAP8) and altered its subcellular localization from the insoluble fraction to the soluble fraction. These results suggest that ALK ATI induces chromatin structural changes and heterochromatinization through phosphorylation of AKAP8 in the nucleus.Key words anaplastic lymphoma kinase; nuclear tyrosine kinase; A-kinase anchoring protein 8; heterochromatinization; alternative transcription initiation; chromatin structural change Chromatin structure and organization are dynamically changed during cellular processes, such as proliferation, differentiation, DNA damage responses, and tumorigenesis.

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Nuclear c-Abl-mediated tyrosine phosphorylation induces chromatin structural changes through histone modifications that include H4K16 hypoacetylation

Cited by 30 publications

References 73 publications

FOXA1 Induces E-Cadherin Expression at the Protein Level <i>via</i> Suppression of Slug in Epithelial Breast Cancer Cells

FOXA1 Induces E-Cadherin Expression at the Protein Level <i>via</i> Suppression of Slug in Epithelial Breast Cancer Cells

Src Acts as an Effector for Ku70-dependent Suppression of Apoptosis through Phosphorylation of Ku70 at Tyr-530

The Truncated Isoform of the Receptor Tyrosine Kinase ALK Generated by Alternative Transcription Initiation (ALK<sup>ATI</sup>) Induces Chromatin Structural Changes in the Nucleus in a Kinase Activity-Dependent Manner

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