Cell Killing by the
            <i>Drosophila</i>
            Gene
            <i>reaper</i>

White, Kristin; Tahaoglu, Elvan; Steller, Hermann

doi:10.1126/science.271.5250.805

Cited by 362 publications

(315 citation statements)

References 18 publications

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confidence: 55%

“…2a, b). Consistent with these expression patterns, when reaper (rpr), an apoptosis-inducible gene (White et al, 1996) was driven by puc GAL4E69 , individuals did not die during the embryonic stage but rather at the early larval stage (data not shown).In the imaginal discs in the third instar larvae, puc GAL4E69 expression was much broader than known puc-lacZ expression (Fig. 2c-j), although the P-element insertion sites in both puc GAL4E69 and puc E69 were identical.…”

mentioning

confidence: 60%

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Puckered‐GAL4 driving in JNK‐active cells

Adachi‐Yamada

2002

Genesis

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c-Jun N-terminal kinase (JNK) is an evolutionarily conserved protein kinase belonging to the mitogen-activated protein kinase (MAPK) superfamily. It regulates a number of cellular processes such as proliferation, differentiation, and apoptosis in response to various growthinducible and stressful stimuli (Davis, 2000). In a genetically amenable organism, fruit fly Drosophila, a homolog of JNK (also known as Basket or DJNK) has been reported to control cell morphogenesis, planar cell polarity, apoptosis, and wound healing (Noselli, 1998;Ip and Davis, 1998;Ramet et al., 2002). To understand the detailed molecular mechanism controlling these phenomena, it is important to examine how modification of signaling affects these cellular responses. The GAL4/ UAS system (Brand and Perrimon, 1993), which functions in JNK-active cells, is a good tool to study in this respect. Here I engineer a GAL4 enhancer trap of the puckered (puc) gene, which encodes a MAPK phosphatase family member to inactivate JNK (Martín-Blanco et al., 1998). The expression of puc is known to be induced by the JNK signal, thereby creating a negative regulatory circuit of JNK signaling. This new GAL4 line was named puc GAL4E69 .A technique that was referred to as enhancer-trap targeting (Gonzy-Treboul et al., 1995) was used to induce the puc GAL4E69 GAL4-enhancer trap line. Briefly, p{A92}, a derivative of the transposon P-element with a reporter gene lacZ, which is included in the puc E69 lacZ-enhancer trap allele, was replaced with pGawB, another P-element construct with an enhancer-less GAL4 gene (Brand and Perrimon, 1993). About 200 dysgenic males possessing three kinds of P-element-i.e. puc E69 , X-linked pGawB, and ⌬2-3, a transposase supplier Pelement-were crossed with normal females, and then approximately 500 male progenies without ⌬2-3 were collected. It is possible that they contained new autosomal insertions. Among this collection, a single line showing a GAL4 expression pattern similar to the known puc-lacZ expression pattern was identified. The structure around the P-element insertion is shown in Figure 1. Sequence analyses revealed that the insertion sites of each puc E69 and puc GAL4E69 P-element were identical. However, the two P-elements p{A92} and pGawB were not precisely replaced. Consequently, the resulting GAL4-expressing construct lost both of the visible dominant markers, ry ϩ in p{A92} and mini-w ϩ in pGawB. It also lost the lacZ reporter gene in p{A92}.The expression pattern of puc GAL4E69 in the embryo is shown in Figure 2a and b. JNK is known to be activated in the leading edge cells of the epithelial cell sheet during embryonic dorsal closure. However, unlike the previously reported puc-lacZ expression (Glise et al., 1995), puc GAL4E69 expression recognized by UAS-lacZ expression could not be detected in the leading edge cells. The failure of this expression was probably caused by the presence of a potential silencer-like element in the newly inserted GAL4-expressing P-element construct. At the later embryonic stage, puc GA...

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supporting

confidence: 55%

mentioning

confidence: 60%

Puckered‐GAL4 driving in JNK‐active cells

Adachi‐Yamada

2002

Genesis

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“…36,37 Drosophila does express two Bcl-2 family members, which are both pro-apoptotic. 3 Their mechanism of action is presently unclear. Expression of mammalian Bcl-2 in Drosophila has yielded contradictory results.…”

Section: Discussionmentioning

confidence: 99%

“…The Drosophila protein Reaper is one of these: it induces apoptosis in both Drosophila and mammalian cells. 3,4 Reaper was originally identified in a genetic screen for molecules involved in developmental cell death in the fruit fly. Loss of H99 chromosomal region, which encodes Reaper, Grim and Hid, results in embryonic cell accumulation and imparts resistance to various apoptotic stimuli.…”

Section: Introductionmentioning

confidence: 99%

Mechanism of action of Drosophila Reaper in mammalian cells: Reaper globally inhibits protein synthesis and induces apoptosis independent of mitochondrial permeability

et al. 2004

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Drosophila Reaper can bind inhibitor of apoptosis proteins (IAP) and thereby rescue caspases from proteasomal degradation. In insect cells, this is sufficient to induce apoptosis. Reaper can also induce apoptosis in mammalian cells, in which caspases need to be activated, usually via the mitochondrial pathway. Nevertheless, we find that Reaper efficiently induces apoptosis in mammalian cells in the absence of mitochondrial permeabilisation and cytochrome c release. Moreover, this capacity was only marginally affected by deletion of Reaper's amino-terminal IAP-binding motif. Independent of this motif, Reaper could globally suppress protein synthesis. Deletion of 20 amino acids from the carboxy-terminus of Reaper fully abrogated its potential to inhibit protein synthesis and to induce apoptosis in the absence of IAP-binding. Our findings indicate that the newly identified capacity of Reaper to suppress protein translation can operate in mammalian cells and may be key to its proapoptotic activity.

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“…10,11 In cells committed to apoptosis, IAP-mediated inhibition of caspases is counteracted by IAP antagonists. Specifically, the IAP antagonists encoded by reaper, hid and grim (RHG) [12][13][14][15] in Drosophila trigger proteolytic degradation of DIAP1 [16][17][18][19][20] which releases caspases from DIAP1 inhibition and triggers apoptosis. The overexpression of the RHG genes in the fly eye using the eye-specific GMR promoter causes an eye ablation phenotype due to massive apoptosis (see, for example, GMR-hid in Figure 1a).…”

mentioning

confidence: 99%

Genetic characterization of two gain-of-function alleles of the effector caspase DrICE in Drosophila

Lindblad

Garnett

et al. 2015

Cell Death Differ

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Caspases are the executioners of apoptosis. Although much is known about their physiological roles and structures, detailed analyses of missense mutations of caspases are lacking. As mutations within caspases are identified in various human diseases, the study of caspase mutants will help to elucidate how caspases interact with other components of the apoptosis pathway and how they may contribute to disease. DrICE is the major effector caspase in Drosophila required for developmental and stressinduced cell death. Here, we report the isolation and characterization of six de novo drICE mutants, all of which carry point mutations affecting amino acids conserved among caspases in various species. These six mutants behave as recessive loss-offunction mutants in a homozygous condition. Surprisingly, however, two of the newly isolated drICE alleles are gain-of-function mutants in a heterozygous condition, although they are loss-of-function mutants homozygously. Interestingly, they only behave as gain-of-function mutants in the presence of an apoptotic signal. These two alleles carry missense mutations affecting conserved amino acids in close proximity to the catalytic cysteine residue. This is the first time that viable gain-of-function alleles of caspases are described in any intact organism and provides a significant exception to the expectation that mutations of conserved amino acids always abolish the pro-apoptotic activity of caspases. We discuss models about how these mutations cause the gain-offunction character of these alleles. Cell Death and Differentiation (2016) 23, 723-732; doi:10.1038/cdd.2015.144; published online 6 November 2015Apoptosis is a major form of programmed cell death. 1 The core apoptotic machinery is evolutionary conserved with caspases as the fundamental components. [1][2][3] Caspases are specific cysteine proteases that are produced as inactive zymogens composed of an N-terminal pro-domain, a large subunit region with the catalytic cysteine residue, and a small subunit region at the carboxyl end. 2,4 Depending on their structures and functions, caspases are grouped into initiator and effector caspases. 2,3 Initiator caspases possess long pro-domains, which facilitate the recruitment of initiator caspases into cell death signaling complexes for activation. 2,3,5 Effector caspases are activated by initiator caspase complexes through proteolytic processing, cleaving off the pro-domain and separating the large and small subunits. The active effector caspase is a tetramer composed of two large and two small subunits and contains two catalytic sites. 2,3 Activated effector caspases cleave many protein targets to trigger the physiological and morphological changes characteristic of apoptosis. In mammals, apoptotic initiator caspases are Caspase-8, -9, -2 and -10, and effector caspases involved in apoptosis are 3 Of the seven Drosophila caspases, only the initiator caspase Dronc (Caspase-9-like) and the effector caspases DrICE and Dcp-1 (Caspase-3-like) have been implicated in apoptosis in imagina...

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Cell Killing by the Drosophila Gene reaper

Cited by 362 publications

References 18 publications

Puckered‐GAL4 driving in JNK‐active cells

Puckered‐GAL4 driving in JNK‐active cells

Mechanism of action of Drosophila Reaper in mammalian cells: Reaper globally inhibits protein synthesis and induces apoptosis independent of mitochondrial permeability

Genetic characterization of two gain-of-function alleles of the effector caspase DrICE in Drosophila

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