Molecular mechanism of staurosporine-induced apoptosis in osteoblasts

Chae, Han‐Jung; Kang, Jian; Byun, Jong-Ook; Han, Kyung-Soo; Kim, Dae-Up; Oh, Sang-Woo; Kim, Hyung-Min; Chae, Soo‐Wan; Kim, Hyung‐Ryong

doi:10.1006/phrs.2000.0700

Cited by 142 publications

(108 citation statements)

References 40 publications

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“…To test whether C. glabrata can cause apoptosis of MDMs, we determined the activity of the effector caspases 3 and 7. As a positive control, MDMs were stimulated with staurosporine, an ATP-competitive protein kinase inhibitor, which is known to induce apoptosis by activating caspase-3 (35). As expected, staurosporine-treated MDMs showed pronounced caspase-3 or caspase-7 activity after 3 h, resulting in a peak of activation after 8 h coincubation (Fig.…”

Section: Glabrata Does Not Induce Macrophage Apoptosissupporting

confidence: 63%

The Facultative Intracellular Pathogen Candida glabrata Subverts Macrophage Cytokine Production and Phagolysosome Maturation

Seider

Brunke

Schild

et al. 2011

The Journal of Immunology

184

226

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Although Candida glabrata is an important human pathogenic yeast, its pathogenicity mechanisms are largely unknown. Immune evasion strategies seem to play key roles during infection, since very little inflammation is observed in mouse models. Furthermore, C. glabrata multiplies intracellularly after engulfment by macrophages. In this study, we sought to identify the strategies that enable C. glabrata to survive phagosome biogenesis and antimicrobial activities within human monocyte-derived macrophages. We show that, despite significant intracellular proliferation, macrophage damage or apoptosis was not apparent, and production of reactive oxygen species was inhibited. Additionally, with the exception of GM-CSF, levels of pro- and anti-inflammatory cytokines were only marginally increased. We demonstrate that adhesion to and internalization by macrophages occur within minutes, and recruitment of endosomal early endosomal Ag 1 and lysosomal-associated membrane protein 1 indicates phagosome maturation. However, phagosomes containing viable C. glabrata, but not heat-killed yeasts, failed to recruit cathepsin D and were only weakly acidified. This inhibition of acidification did not require fungal viability, but it had a heat-sensitive surface attribute. Therefore, C. glabrata modifies the phagosome into a nonacidified environment and multiplies until the host cells finally lyse and release the fungi. Our results suggest persistence of C. glabrata within macrophages as a possible immune evasion strategy.

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Section: Glabrata Does Not Induce Macrophage Apoptosissupporting

confidence: 63%

The Facultative Intracellular Pathogen Candida glabrata Subverts Macrophage Cytokine Production and Phagolysosome Maturation

Seider

Brunke

Schild

et al. 2011

The Journal of Immunology

184

226

View full text Add to dashboard Cite

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“…Jurkat cells were infected with WT VACV (VACV-EGFP), VACV devoid of F1L (VACV⌬F1L), or VACV expressing Flag-tagged F1L binding pocket mutations. To further promote Bak activation, mock-infected and infected cells were treated in the presence or absence of STS, a potent inducer of intrinsic apoptosis (50). Numerous viruses were able to completely prevent Bak activation, including VACV-EGFP (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…F1L binding pocket mutants with binding phenotypes similar to those of WT F1L were fully capable of preventing Bak activation. Furthermore, these mutants were able to prevent cytochrome c release from the mitochondria and were capable of preventing the downstream cleavage of the apoptotic indicator PARP (2,50). Binding cleft mutants whose ability to bind both Bim L and Bak was abrogated were dramatically impaired in these assays, including both F1L(N136F) and F1L(F148A) ( Table 2).…”

Section: Discussionmentioning

confidence: 99%

Structural Insight into BH3 Domain Binding of Vaccinia Virus Antiapoptotic F1L

Campbell

Thibault

Mehta

et al. 2014

J Virol

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Apoptosis is a tightly regulated process that plays a crucial role in the removal of virus-infected cells, a process controlled by both pro-and antiapoptotic members of the Bcl-2 family. The proapoptotic proteins Bak and Bax are regulated by antiapoptotic Bcl-2 proteins and are also activated by a subset of proteins known as BH3-only proteins that perform dual functions by directly activating Bak and Bax or by sequestering and neutralizing antiapoptotic family members. Numerous viruses express proteins that prevent premature host cell apoptosis. Vaccinia virus encodes F1L, an antiapoptotic protein essential for survival of infected cells that bears no discernible sequence homology to mammalian cell death inhibitors. Despite the limited sequence similarities, F1L has been shown to adopt a novel dimeric Bcl-2-like fold that enables hetero-oligomeric binding to both Bak and the proapoptotic BH3-only protein Bim that ultimately prevents Bak and Bax homo-oligomerization. However, no structural data on the mode of engagement of F1L and its Bcl-2 counterparts are available. Here we solved the crystal structures of F1L in complex with two ligands, Bim and Bak. Our structures indicate that F1L can engage two BH3 ligands simultaneously via the canonical Bcl-2 ligand binding grooves. Furthermore, by structure-guided mutagenesis, we generated point mutations within the binding pocket of F1L in order to elucidate the residues responsible for both Bim and Bak binding and prevention of apoptosis. We propose that the sequestration of Bim by F1L is primarily responsible for preventing apoptosis during vaccinia virus infection. IMPORTANCENumerous viruses have adapted strategies to counteract apoptosis by encoding proteins responsible for sequestering proapoptotic components. Vaccinia virus, the prototypical member of the family Orthopoxviridae, encodes a protein known as F1L that functions to prevent apoptosis by interacting with Bak and the BH3-only protein Bim. Despite recent structural advances, little is known regarding the mechanics of binding between F1L and the proapoptotic Bcl-2 family members. Utilizing three-dimensional structures of F1L bound to host proapoptotic proteins, we generated variants of F1L that neutralize Bim and/or Bak. We demonstrate that during vaccinia virus infection, engagement of Bim and Bak by F1L is crucial for subversion of host cell apoptosis.

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“…14,16 In particular, we examined the effects on cell viability of four types of stressors: (i) buthionine sulphoximine (BSO), which decreases intracellular glutathione by inhibition of gammaglutamylcysteine synthetase; 24 (ii) staurosporine, a kinase inhibitor that induces apoptotic cell death; 25 (iii) paraquat, a superoxide generator; and (iv) hydrogen peroxide. Each stressor was tested at a range of doses and times as used in previous studies 14,16,24,25 for the effect on cell viability (Supplementary Methods).…”

Section: Stressor Responses In Cultures Of Fshd and Unaffected Contromentioning

confidence: 99%

A unique library of myogenic cells from facioscapulohumeral muscular dystrophy subjects and unaffected relatives: family, disease and cell function

et al. 2011

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To explore possible mechanisms of pathology in facioscapulohumeral muscular dystrophy (FSHD), we generated a novel library of myogenic cells composed of paired cultures derived from FSHD subjects and unaffected first-degree relatives. We prepared cells from biopsies of both biceps and deltoid muscles obtained from each of 10 FSHD and 9 unaffected donors. We used this new collection to determine how family background and disease affected patterns of growth and differentiation, expression of a panel of candidate, and muscle-specific genes, and responses to exogenous stressors. We found that FSHD and unaffected cells had, on average, indistinguishable patterns of differentiation, gene expression, and dose-response curves to staurosporine, paraquat, hydrogen peroxide, and glutathione depletion. Differentiated FSHD and unaffected cultures were both more sensitive to glutathione depletion than proliferating cultures, but showed similar responses to paraquat, staurosporine, and peroxide. For stress responses, the sample size was sufficient to detect a 10% change in effect at the observed variability with a power of 499%. In contrast, for each of these properties, we found significant differences among cells from different cohorts, and these differences were independent of disease status, gender, or muscle biopsied. Thus, though none of the properties we examined could be used to reliably distinguish between FSHD and unaffected cells, family of origin was an important contributor to gene-expression patterns and stressor responses in cultures of both FSHD and unaffected myogenic cells.

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Molecular mechanism of staurosporine-induced apoptosis in osteoblasts

Cited by 142 publications

References 40 publications

The Facultative Intracellular Pathogen Candida glabrata Subverts Macrophage Cytokine Production and Phagolysosome Maturation

The Facultative Intracellular Pathogen Candida glabrata Subverts Macrophage Cytokine Production and Phagolysosome Maturation

Structural Insight into BH3 Domain Binding of Vaccinia Virus Antiapoptotic F1L

A unique library of myogenic cells from facioscapulohumeral muscular dystrophy subjects and unaffected relatives: family, disease and cell function

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