Caspase-catalyzed cleavage and activation of Mst1 correlates with eosinophil but not neutrophil apoptosis

Souza, Patrícia Mendes de; Kankaanranta, Hannu; Michael, Ashour; Barnes, Peter J.; Giembycz, Mark A.; Lindsay, Mark A.

doi:10.1182/blood.v99.9.3432

Cited by 43 publications

(32 citation statements)

References 28 publications

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“…[7][8][9] Furthermore, 36-kDa Mst1 is activated during apoptosis by anti-Fas antibody in human eosinophils and CD4 ϩ T-lymphocytes, but not in human neutrophils. 12,13) In the present study, activation of 36-kDa MBP kinase by antiFas antibody, dexamethasone and aminophylline also paralleled our observation of histone H2B phosphorylation. Cheung et al 10) demonstrated that catalytic 36-kDa Mst1 phosphorylated histone H2B in HL-60 cells.…”

Section: Discussionsupporting

confidence: 75%

“…12,13) In the present study, we investigated the effect of dexamethasone and aminophylline (a theophylline dimer) on survival of T-lymphocytic leukemia Jurkat and promyelocytic leukemia HL-60 cells.…”

Section: -9)mentioning

confidence: 99%

“…10,11) The promotion of apoptosis of eosinophils and CD4 ϩ T cells, but not neutrophils, is reportedly associated with the activation of 36-kDa Mst1. 12,13) In the present study, we investigated the effect of dexamethasone and aminophylline (a theophylline dimer) on survival of T-lymphocytic leukemia Jurkat and promyelocytic leukemia HL-60 cells.…”

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confidence: 99%

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Effects of Dexamethasone and Aminophylline on Survival of Jurkat and HL-60 Cells

Kizawa

Furuya

Saito

et al. 2006

Biological & Pharmaceutical Bulletin

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Glucocorticoids are currently the most potent anti-inflammatory drugs available and, whether administered systemically or by inhalation, are first-line drugs for the treatment of many respiratory diseases such as asthma. Theophylline is also known to have an anti-inflammatory action that may account for its clinical effectiveness in the reduction of inflammatory cells in the airway, in addition to its bronchodilative effects. There is increasing evidence that not only glucocorticoid, but also theophylline, attenuates eosinophil accumulation in the airways of asthmatic patients. 1-3)Glucocorticoids and theophylline shorten the half-life of some inflammatory cells such as eosinophils. The survival of these cells depends on the presence of cytokines such as interleukin 5 and GM-CSF. Glucocorticoids and theophylline block the synthesis of these cytokines and so trigger apoptosis, but the molecular mechanism of this effect has yet to be elucidated. [4][5][6] Several studies have shown that Mst1 (mammalian ste20-like kinase 1), a group II germinal center kinase, generates a catalytically active 36-kDa fragment during apoptosis. 7-9)The 36-kDa Mst1 has been reported to catalyse the phosphorylation of histone H2B at Ser 14 , which is associated with chromosome condensation during apoptosis.10,11) The promotion of apoptosis of eosinophils and CD4 ϩ T cells, but not neutrophils, is reportedly associated with the activation of 36-kDa Mst1. 12,13) In the present study, we investigated the effect of dexamethasone and aminophylline (a theophylline dimer) on survival of T-lymphocytic leukemia Jurkat and promyelocytic leukemia HL-60 cells. MATERIALS AND METHODSCell Culture Jurkat (clone E6-1, ATCC, VA, U.S.A.) and HL-60 (clone 15, ATCC, VA, U.S.A.) cells were cultured in RPMI 1640 medium supplemented with 10% FBS and 2 mM L-glutamine in a 5% CO 2 incubator at 37°C. The medium was refreshed every 2 or 3 d. HL-60 (clone 15) cell line is known to differentiate eosinophils, neutrophils and monocytes.Determination of Apoptosis Apoptotic cells were quantified using Annexin V flow cytometry as described by Vermes et al.14) The Annexin V assay exploits the fact that an early event during apoptosis of many cells is a loss of membrane lipid asymmetry, resulting in exposure of phosphatidylserine in the outer leaflet of the plasma membrane. Briefly, cells were incubated with FITC-conjugated Annexin V and counterstained with propidium iodide (PI) in order to allow exclusion of necrotic cells. The cells were subsequently analyzed using a flow cytometer (Beckman Coulter Japan, Tokyo, Japan).Detection of Phospho-Histone H2B (Ser 14 )-Positive Cells Cells grown on chamber slides (Asahi Technoglass) were fixed in 4% paraformaldehyde and then washed in PBS. Following permeabilization with 0.2% Triton X-100, cells were blocked in 2% goat serum PBS (blocking solution), then incubated with the H2B S14 phospho antibodies in block solution at 37°C. The cells were then washed three times with blocking solution and incubated for 1 h in goat anti-rabbit ...

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Section: Discussionsupporting

confidence: 75%

Section: -9)mentioning

confidence: 99%

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Effects of Dexamethasone and Aminophylline on Survival of Jurkat and HL-60 Cells

Kizawa

Furuya

Saito

et al. 2006

Biological & Pharmaceutical Bulletin

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“…6B, lane 3), the mass of cyclin D3 is comparable to that seen in lane 1. Inclusion of a caspase inhibitor, known to effectively increase eosinophil survival in the absence of added cytokine (100 M Z-Asp-CH 2 -[(2,6-dichlorobenzoyl)oxy]methane) (33), did not attenuate the decrease in cyclin D3 protein levels (Fig. 6B, lane 4), whereas an inhibitor of the ubiquitin proteosome system, MG115 (50 M), prevented the loss of immunoreactive cyclin D3 (Fig.…”

Section: Regulation Of Cyclin D3 Protein Expression In Eosinophils Bymentioning

confidence: 99%

IL-5 and Granulocyte-Macrophage Colony-Stimulating Factor Activate STAT3 and STAT5 and Promote Pim-1 and Cyclin D3 Protein Expression in Human Eosinophils

Stout

Bates

Liu

et al. 2004

The Journal of Immunology

109

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Allergic inflammation is characterized by elevated eosinophil numbers and by the increased production of the cytokines IL-5 and GM-CSF, which control several eosinophil functions, including the suppression of apoptosis. The JAK/STAT pathway is important for several functions in hemopoietic cells, including the suppression of apoptosis. We report in this study that STAT3, STAT5a, and STAT5b are expressed in human eosinophils and that their signaling pathways are active following IL-5 or GM-CSF treatment. However, in airway eosinophils, the phosphorylation of STAT5 by IL-5 is reduced, an event that may be related to the reduced expression of the IL-5Rα on airway eosinophils. Furthermore, IL-5 and GM-CSF induced the protein expression of cyclin D3 and the kinase Pim-1, both of which are regulated by STAT-dependent processes in some cell systems. Pim-1 is more abundantly expressed in airway eosinophils than in blood eosinophils. Because Pim-1 reportedly has a role in the modulation of apoptosis, these results suggest that Pim-1 action is linked to the suppression of eosinophil apoptosis by these cytokines. Although cyclin D3 is known to be critical for cell cycle progression, eosinophils are terminally differentiated cells that do not proceed through the cell cycle. Thus, this apparent cytokine regulation of cyclin D3 suggests that there is an alternative role(s) for cyclin D3 in eosinophil biology.

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“…MST1, for example, can induce apoptosis and nuclear condensation in BJAB, 293T, and COS-1 cells (15,46,47), and MST1 activity correlates with eosinophil apoptosis (10). Expression of MST3 in HEK293 cells leads to DNA fragmentation and apoptosis (19).…”

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confidence: 99%

Regulation of NDR Protein Kinase by Hydrophobic Motif Phosphorylation Mediated by the Mammalian Ste20-Like Kinase MST3

Stegert

Hergovich

Tamaskovic

et al. 2005

Molecular and Cellular Biology

141

163

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NDR protein kinases are involved in the regulation of cell cycle progression and morphology. NDR1/NDR2 protein kinase is activated by phosphorylation on the activation loop phosphorylation site Ser281/Ser282 and the hydrophobic motif phosphorylation site Thr444/Thr442. Autophosphorylation of NDR is responsible for phosphorylation on Ser281/Ser282, whereas Thr444/Thr442 is targeted by an upstream kinase. Here we show that MST3, a mammalian Ste20-like protein kinase, is able to phosphorylate NDR protein kinase at Thr444/ Thr442. In vitro, MST3 selectively phosphorylated Thr442 of NDR2, resulting in a 10-fold stimulation of NDR activity. MOB1A (Mps one binder 1A) protein further increased the activity, leading to a fully active kinase. In vivo, Thr442 phosphorylation after okadaic acid stimulation was potently inhibited by MST3KR, a kinasedead mutant of MST3. Knockdown of MST3 using short hairpin constructs abolished Thr442 hydrophobic motif phosphorylation of NDR in HEK293F cells. We conclude that activation of NDR is a multistep process involving phosphorylation of the hydrophobic motif site Thr444/2 by MST3, autophosphorylation of Ser281/2, and binding of MOB1A.The NDR and LATS family of serine/threonine protein kinases participates in the regulation of cell cycle progression and cell morphology (14,43,53). These kinases share a conserved N-terminal regulatory domain that interacts with MOB (3, 6, 18, 33, 50) and S100B (31) proteins. The conserved catalytic (kinase) domain has an insertion of 30 amino acids between subdomains VII and VIII containing the activation segment phosphorylation site Ser281/Ser282 and, in the case of mammalian NDR1 and NDR2, an autoinhibitory sequence (3). The C-terminal regulatory domain encompasses the regulatory hydrophobic motif phosphorylation site Thr444 (32). Recent data show that NDR1 and NDR2 protein kinase activities are stimulated by Ca 2ϩ /S100B-and MOB1-binding-induced autophosphorylation on S281/S282 in vitro and in vivo (3, 31). Phosphorylation of the hydrophobic motif phosphorylation site T444/T442 is required for maximal activation and involves an upstream kinase (42,44).Genetic evidence suggests that Ste20 (Sterile)-like protein kinases function as upstream kinases of the NDR family. For example, one of the Saccharomyces cerevisiae Ste20-like kinases, Cdc15p, phosphorylates Dbf2p (28), and the Schizosaccharomyces pombe Ste20-like kinase Pak1p/Shk1p genetically interacts with Orb6p (49). Furthermore, the Ste20-like kinase Kic1p functionally interacts with Cbk1p, the closest relative of NDR from Saccharomyces cerevisiae (33), and the Drosophila melanogaster Ste20-like kinase HIPPO phosphorylates WARTS/LATS kinase (16,20,34,45,51). The closest mammalian homologues of Kic1p and Hippo are the mammalian Ste20-like protein kinases MST1/KRS1, MST2/KRS2, MST3, MST4/MASK, and SOK/YSK (29), which are involved in the regulation of cell morphology, proliferation, and apoptosis (7,9,22,25,38). This group of kinases contains an N-terminal kinase domain as well as an autoinhibitory C-...

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Caspase-catalyzed cleavage and activation of Mst1 correlates with eosinophil but not neutrophil apoptosis

Cited by 43 publications

References 28 publications

Effects of Dexamethasone and Aminophylline on Survival of Jurkat and HL-60 Cells

Effects of Dexamethasone and Aminophylline on Survival of Jurkat and HL-60 Cells

IL-5 and Granulocyte-Macrophage Colony-Stimulating Factor Activate STAT3 and STAT5 and Promote Pim-1 and Cyclin D3 Protein Expression in Human Eosinophils

Regulation of NDR Protein Kinase by Hydrophobic Motif Phosphorylation Mediated by the Mammalian Ste20-Like Kinase MST3

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