Caspase-9 CARD : core domain interactions require a properly formed active site

Huber, Kristen L.; Serrano, Banyuhay P.; Hardy, James L.

doi:10.1042/bcj20170913

Cited by 20 publications

(26 citation statements)

References 63 publications

(86 reference statements)

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“…24 Caspase-9 is the upstream caspase which is involved in the apoptotic process. [25][26][27] In the current study, we observed that the activities of caspase-3 and caspase-9 proteins induced, these findings suggested that Res increased the apoptosis of HSC-3 cells via increasing the activities of caspase-3 and caspase-9.…”

Section: Discussionsupporting

confidence: 64%

Resveratrol inhibits oral squamous cell carcinoma cells proliferation while promoting apoptosis through inhibition of CBX7 protein

Chen

Xia

et al. 2020

Environmental Toxicology

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As a natural compound, resveratrol (Res) is confirmed to be promising drug for the treatment of malignant tumors. Therefore, our study aimed to observe the impacts of Res on the proliferation and apoptosis of oral squamous cell carcinoma cells (HSC-3 cells) as well as the mechanism involving chromobox protein homolog 7 (CBX7) signal transduction. HSC-3 cells were treated with Res, Akt agonist (AL3818) and p16 inhibitor (SC79), and transfected with CBX7 mimics and inhibitor plasmids. The CCK-8 assay was used to detect cell proliferation, flow cytometry was performed to assess cell cycle and apoptosis, and cell colonies and histone DNA level were also measured. Western blot analysis was used to determine the expression levels of related proteins. HSC-3 cells showed decreased cell proliferation, colonies, BrdU-counled cells and increased apoptosis, histone DNA level, the activities of caspase-3 and caspase-9 when treated with Res. Western blot analysis revealed elevated Cle-PARP and Cle-caspase 3 expression and reduced t-PARP expression in HSC-3 cells treated with Res compared with control. AL3818 and SC79 could decrease the inhibitory effects of Res on the growth of HSC-3 cells. Furthermore, CBX7 overexpression could also partly reverse the roles of Res in the growth of HSC-3 cells, and Akt and p16 signal transduction. Our results demonstrate that Res suppresses the proliferation, and induces the apoptosis of oral squamous cell carcinoma cells through the inhibition of CBX7/Akt and the activation of p16 cascades.

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Section: Discussionsupporting

confidence: 64%

Resveratrol inhibits oral squamous cell carcinoma cells proliferation while promoting apoptosis through inhibition of CBX7 protein

Chen

Xia

et al. 2020

Environmental Toxicology

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“…5b and Supplementary Fig. 5) 60 . We then used wildtype caspase-9 monomer C and inactive monomer I to assemble homo-and heterodimeric DNA nanostructures, and confirmed that the heterodimeric variants assembled correctly using agarose gel electrophoresis ( Supplementary Fig.…”

Section: Resultsmentioning

confidence: 98%

A DNA-based synthetic apoptosome

Bj¹,

Aj²,

Gumí-Audenis³

et al. 2019

Preprint

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Living cells are able to regulate key cellular processes by physically assembling signaling components on dedicated molecular platforms. The spatial organization of proteins in these higher-order signaling complexes facilitates proximity-driven activation and inhibition events, allowing tight regulation of the flow of information. Here, we employ the programmability and modularity of DNA origami as a controllable molecular platform for studying protein-protein interactions involved in intracellular signaling. Specifically, we engineer a synthetic, DNA origami-based version of the apoptosome, a large multi-protein signaling complex that regulates apoptosis by co-localization of multiple caspase-9 monomers. Our in vitro characterization using both wildtype caspase-9 monomers and inactive mutants tethered to a DNA origami platform directly demonstrates that enzymatic activity is induced by proximity-driven dimerization with asymmetric, half-of-sites reactivity. Additionally, experimental results supported by a detailed thermodynamic model reveal a multivalent activity enhancement in tethered caspase-9 oligomers of three and four enzymes, partly originating from a statistical increase in the number of active catalytic units in higherorder enzyme clusters. Our results offer fundamental insights in caspase-9 activity regulation and demonstrate that DNA origami provides a modular platform to construct and characterize higher-order signaling complexes. The engineered DNA-based protein assembly platform has the potential to be broadly applied to inform the function of other important multi-enzyme assemblies involved in inflammation, innate immunity, and necrosis.

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“…As such, several features of the native system that could modulate caspase-9 activation and regulation are not taken into account. For example, recent work has indicated that allosteric interactions of the catalytic domain of caspase-9 with its CARD, the flexible linkers, and the apoptosome itself can affect enzyme activity 36,53,54 . However, we argue that a modular DNA-based model system composed of a controlled number of catalytic domains with well-defined interactions can help isolate the key molecular determinants of caspase-9 regulation.…”

Section: Dna Origami-mediated Caspase-9 Enzyme Activitymentioning

confidence: 99%

Proximity-induced caspase-9 activation on a DNA origami-based synthetic apoptosome

et al. 2020

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Living cells regulate key cellular processes by spatial organisation of catalytically active proteins in higher-order signalling complexes. These act as organising centres to facilitate proximityinduced activation and inhibition of multiple intrinsically weakly associating signalling components, which makes elucidation of the underlying protein-protein interactions challenging. Here we show that DNA origami nanostructures provide a programmable molecular platform for the systematic analysis of signalling proteins by engineering a synthetic DNA origami-based version of the apoptosome, a multi-protein complex that regulates apoptosis by co-localizing multiple caspase-9 monomers. Tethering of both wildtype and inactive caspase-9 variants to a DNA origami platform demonstrates that enzymatic activity is induced by proximity-driven dimerization with half-of-sites reactivity, and additionally, reveals a multivalent activity enhancement in oligomers of three and four enzymes. Our results offer fundamental insights in Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:

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Caspase-9 CARD : core domain interactions require a properly formed active site

Cited by 20 publications

References 63 publications

Resveratrol inhibits oral squamous cell carcinoma cells proliferation while promoting apoptosis through inhibition of CBX7 protein

Resveratrol inhibits oral squamous cell carcinoma cells proliferation while promoting apoptosis through inhibition of CBX7 protein

A DNA-based synthetic apoptosome

Proximity-induced caspase-9 activation on a DNA origami-based synthetic apoptosome

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