Functional Interplay between Caspase Cleavage and Phosphorylation Sculpts the Apoptotic Proteome

Dix, Melissa M.; Simón, Gabriel; Wang, Chu; Okerberg, Eric; Patricelli, Matthew P.; Cravatt, Benjamin F.

doi:10.1016/j.cell.2012.05.040

Cited by 133 publications

(117 citation statements)

References 52 publications

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“…In addition, we envision the approach to be beneficial in characterizing the cross-talk between phosphorylation and apoptotic proteolysis by including either phosphorylation mimetic or nonphosphorylatable residues in the TevS allele series (40,41). The approach outlined with the PTGR and SNIPer technology may prove particularly beneficial in profiling site-specific proteolysis throughout cell biology: for example, during cell-cycle progression mediated by separase during anaphase (42), differentiation, and intercellular signaling by Notch proteolysis (43), and mediating the unfolded protein response by ATF6 proteolysis (44).…”

Section: Discussionmentioning

confidence: 99%

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

Morgan

Diaz

Zeitlin

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Cellular demolition during apoptosis is completed by executioner caspases, that selectively cleave more than 1,500 proteins but whose individual roles are challenging to assess. Here, we used an optimized site-specific and inducible protease to examine the role of a classic apoptotic node, the caspase-activated DNase (CAD). CAD is activated when caspases cleave its endogenous inhibitor ICAD, resulting in the characteristic DNA laddering of apoptosis. We describe a posttranscriptional gene replacement (PTGR) approach where endogenous biallelic ICAD is knocked down and simultaneously replaced with an engineered allele that is susceptible to inducible cleavage by tobacco etch virus protease. Remarkably, selective activation of CAD alone does not induce cell death, although hallmarks of DNA damage are detected in human cancer cell lines. Our data strongly support that the highly cooperative action of CAD and inhibition of DNA repair systems are critical for the DNA laddering phenotype in apoptosis. Furthermore, the PTGR approach provides a general means for replacing wild-type protein function with a precisely engineered mutant at the transcriptional level that should be useful for cell engineering studies.DNA damage | apoptosis | ICAD | site-specific proteolysis |

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Section: Discussionmentioning

confidence: 99%

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

Morgan

Diaz

Zeitlin

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…In looking for processes capable of causing large-scale changes in physicochemical properties of the cytosol, activation of proteolytic enzymes is the natural candidate. Indeed, 5-10% of all proteins become cleaved during apoptosis (29,107), which may result in an increased exposure of hydrophobic residues. Major changes in the cytoskeleton that develop in apoptotic cells (17,88) may also affect water structure.…”

Section: Osmotically Inactive Volumementioning

confidence: 99%

Possible causes of apoptotic volume decrease: an attempt at quantitative review

Model

2014

American Journal of Physiology-Cell Physiology

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Model MA. Possible causes of apoptotic volume decrease: an attempt at quantitative review. Am J Physiol Cell Physiol 306: C417-C424, 2014. First published November 6, 2013; doi:10.1152/ajpcell.00328.2013.-Cell shrinkage and dehydration are essential characteristics of apoptosis, and loss of as much as half of the initial cell volume is not uncommon. This phenomenon is usually explained by efflux of K ϩ and Cl Ϫ . We reexamine this hypothesis on the basis of the available data for ion concentrations and the requirements for osmotic equilibrium and electroneutrality. In addition to ion loss, we discuss the possible impacts of several other processes: efflux of low-molecular-weight osmolytes, acidification of the cytosol, effects of water channels and pumps, heterogeneity of intracellular water, and dissociation of apoptotic bodies. We conclude that most mammalian cells are theoretically capable of reducing their volume by 15-20% through ion loss or a decrease in cytosolic pH, although, in reality, the contribution of these mechanisms to apoptotic shrinkage may be smaller. Transitions between osmotically active and inactive water pools might influence cell volume as well; these mechanisms are poorly understood but are amenable to experimental study. Dissociation of apoptotic bodies is a separate mechanism of volume reduction and should be monitored closely; this can be best achieved by measurement of intracellular water, rather than cell volume. cell volume regulation; apoptosis; apoptotic volume decrease; intracellular water CELL SHRINKAGE is one the most reproducible signs of apoptosis (33,103). It has at least three distinct aspects: 1) weakening of the substrate attachment, resulting in cell rounding and reduced spreading, 2) dissociation of cell fragments known as apoptotic bodies, and 3) loss of intracellular water. Apoptotic body formation and the loss of water bring about a decrease in total cell volume. Volume changes in apoptotic cells have been measured by various techniques, and shrinkage by as much as 40 -85% has been reported (11,16,34,53,76,90). Shrinkage due to dehydration has received the most attention in the literature and is sometimes referred to as "apoptotic volume decrease" (AVD).The place of AVD in the sequence of apoptotic events and its significance for the rest of apoptosis have been addressed in numerous research papers and reviews (46,70,71,92,94). Apparently, AVD can be an early (5,8,15,28,34,49,53,81) or a late (26,60,85,90,117) response, depending on the particular system. A few exceptions to the rule that AVD must accompany apoptosis have also been described (11,13,56,65,121,127); nevertheless, it appears that, in the vast majority of apoptotic models, AVD does occur sooner or later.It has been hypothesized that the main purpose of AVD is to facilitate detachment of apoptotic bodies (91) or to oppose cell swelling and rupture that might develop otherwise (128). Although the definitive answer to this question is lacking, there can be little doubt that such a highly conserved property...

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“…Although many individual caspase substrates have been implicated in specific aspects of cellular destruction (e.g., lamin cleavage is required for the efficient packaging of nuclei into small membrane-bound vesicles), recent proteomic approaches have greatly expanded the known repertoire of proteolytic products generated during apoptosis (Van Damme et al 2005;Dix et al 2008;Mahrus et al 2008). Further work will be needed to confirm these findings and to determine how (or if ) all of these substrates participate in the apoptotic process (see Poreba et al 2013), especially as new details emerge on the relationship between posttranslational modifications, like phosphorylation, and caspase cleavage (Dix et al 2012).…”

mentioning

confidence: 99%

Cellular Mechanisms Controlling Caspase Activation and Function

Parrish

Freel

Kornbluth

2013

Cold Spring Harbor Perspectives in Biology

513

367

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Caspases are the primary drivers of apoptotic cell death, cleaving cellular proteins that are critical for dismantling the dying cell. Initially translated as inactive zymogenic precursors, caspases are activated in response to a variety of cell death stimuli. In addition to factors required for their direct activation (e.g., dimerizing adaptor proteins in the case of initiator caspases that lie at the apex of apoptotic signaling cascades), caspases are regulated by a variety of cellular factors in a myriad of physiological and pathological settings. For example, caspases may be modified posttranslationally (e.g., by phosphorylation or ubiquitylation) or through interaction of modulatory factors with either the zymogenic or active form of a caspase, altering its activation and/or activity. These regulatory events may inhibit or enhance enzymatic activity or may affect activity toward particular cellular substrates. Finally, there is emerging literature to suggest that caspases can participate in a variety of cellular processes unrelated to apoptotic cell death. In these settings, it is particularly important that caspases are maintained under stringent control to avoid inadvertent cell death. It is likely that continued examination of these processes will reveal new mechanisms of caspase regulation with implications well beyond control of apoptotic cell death.

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Functional Interplay between Caspase Cleavage and Phosphorylation Sculpts the Apoptotic Proteome

Cited by 133 publications

References 52 publications

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

Possible causes of apoptotic volume decrease: an attempt at quantitative review

Cellular Mechanisms Controlling Caspase Activation and Function

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