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Debcl, a proapoptotic Bcl-2 homologue, is a component of the Drosophila melanogaster cell death machinery.

Colussi PA, Quinn LM, Huang DC, Coombe M, Read SH, Richardson H, Kumar S - J. Cell Biol. (2000)

Bottom Line: Both proapoptotic and antiapoptotic members of this family are found in mammalian cells, but no such proteins have been described in insects.RNA interference studies indicate that Debcl is required for developmental apoptosis in Drosophila embryos.These results suggest that the main components of the mammalian apoptosis machinery are conserved in insects.

View Article: PubMed Central - PubMed

Affiliation: The Hanson Centre for Cancer Research, Institute of Medical and Veterinary Science, Adelaide, SA 5000, Australia.

ABSTRACT
Bcl-2 family of proteins are key regulators of apoptosis. Both proapoptotic and antiapoptotic members of this family are found in mammalian cells, but no such proteins have been described in insects. Here, we report the identification and characterization of Debcl, the first Bcl-2 homologue in Drosophila melanogaster. Structurally, Debcl is similar to Bax-like proapoptotic Bcl-2 family members. Ectopic expression of Debcl in cultured cells and in transgenic flies causes apoptosis, which is inhibited by coexpression of the baculovirus caspase inhibitor P35, indicating that Debcl is a proapoptotic protein that functions in a caspase-dependent manner. debcl expression correlates with developmental cell death in specific Drosophila tissues. We also show that debcl genetically interacts with diap1 and dark, and that debcl-mediated apoptosis is not affected by gene dosage of rpr, hid, and grim. Biochemically, Debcl can interact with several mammalian and viral prosurvival Bcl-2 family members, but not with the proapoptotic members, suggesting that it may regulate apoptosis by antagonizing prosurvival Bcl-2 proteins. RNA interference studies indicate that Debcl is required for developmental apoptosis in Drosophila embryos. These results suggest that the main components of the mammalian apoptosis machinery are conserved in insects.

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debcl mRNA expression in Drosophila. A, Northern blot of poly A+-enriched RNA isolated from various developmental stages and adult flies. debcl transcript is detected as a single, ∼1.5-kb band in most samples examined. The lower panels depict portions of the ethidium bromide-stained gels corresponding to the residual ribosomal RNA bands before transfer to membrane. B, RT-PCR analysis of debcl expression. After reverse transcription of RNA from various stages of Drosophila development, PCR was carried out for 30 cycles using debcl-specific primers that generate a 450-bp product. Lanes 1–4 in B correspond to lanes 1–4 in A. Note that all samples express debcl transcript.
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Figure 2: debcl mRNA expression in Drosophila. A, Northern blot of poly A+-enriched RNA isolated from various developmental stages and adult flies. debcl transcript is detected as a single, ∼1.5-kb band in most samples examined. The lower panels depict portions of the ethidium bromide-stained gels corresponding to the residual ribosomal RNA bands before transfer to membrane. B, RT-PCR analysis of debcl expression. After reverse transcription of RNA from various stages of Drosophila development, PCR was carried out for 30 cycles using debcl-specific primers that generate a 450-bp product. Lanes 1–4 in B correspond to lanes 1–4 in A. Note that all samples express debcl transcript.

Mentions: debcl expression was determined by RNA blotting, RT-PCR, and in situ hybridization to Drosophila embryos and tissues. In most cases low levels of debcl mRNA expression was detected. In RNA blots, a 1.5-kb transcript was evident in most developmental stages, but expression was relatively high in 0–4-h embryos and adult female flies (Fig. 2 A). In late embryos, larvae and pupae debcl expression was somewhat reduced and barely detectable by Northern blotting. However, RT-PCR analysis indicated that debcl mRNA was present in all developmental stages examined (Fig. 2 B).


Debcl, a proapoptotic Bcl-2 homologue, is a component of the Drosophila melanogaster cell death machinery.

Colussi PA, Quinn LM, Huang DC, Coombe M, Read SH, Richardson H, Kumar S - J. Cell Biol. (2000)

debcl mRNA expression in Drosophila. A, Northern blot of poly A+-enriched RNA isolated from various developmental stages and adult flies. debcl transcript is detected as a single, ∼1.5-kb band in most samples examined. The lower panels depict portions of the ethidium bromide-stained gels corresponding to the residual ribosomal RNA bands before transfer to membrane. B, RT-PCR analysis of debcl expression. After reverse transcription of RNA from various stages of Drosophila development, PCR was carried out for 30 cycles using debcl-specific primers that generate a 450-bp product. Lanes 1–4 in B correspond to lanes 1–4 in A. Note that all samples express debcl transcript.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2169366&req=5

Figure 2: debcl mRNA expression in Drosophila. A, Northern blot of poly A+-enriched RNA isolated from various developmental stages and adult flies. debcl transcript is detected as a single, ∼1.5-kb band in most samples examined. The lower panels depict portions of the ethidium bromide-stained gels corresponding to the residual ribosomal RNA bands before transfer to membrane. B, RT-PCR analysis of debcl expression. After reverse transcription of RNA from various stages of Drosophila development, PCR was carried out for 30 cycles using debcl-specific primers that generate a 450-bp product. Lanes 1–4 in B correspond to lanes 1–4 in A. Note that all samples express debcl transcript.
Mentions: debcl expression was determined by RNA blotting, RT-PCR, and in situ hybridization to Drosophila embryos and tissues. In most cases low levels of debcl mRNA expression was detected. In RNA blots, a 1.5-kb transcript was evident in most developmental stages, but expression was relatively high in 0–4-h embryos and adult female flies (Fig. 2 A). In late embryos, larvae and pupae debcl expression was somewhat reduced and barely detectable by Northern blotting. However, RT-PCR analysis indicated that debcl mRNA was present in all developmental stages examined (Fig. 2 B).

Bottom Line: Both proapoptotic and antiapoptotic members of this family are found in mammalian cells, but no such proteins have been described in insects.RNA interference studies indicate that Debcl is required for developmental apoptosis in Drosophila embryos.These results suggest that the main components of the mammalian apoptosis machinery are conserved in insects.

View Article: PubMed Central - PubMed

Affiliation: The Hanson Centre for Cancer Research, Institute of Medical and Veterinary Science, Adelaide, SA 5000, Australia.

ABSTRACT
Bcl-2 family of proteins are key regulators of apoptosis. Both proapoptotic and antiapoptotic members of this family are found in mammalian cells, but no such proteins have been described in insects. Here, we report the identification and characterization of Debcl, the first Bcl-2 homologue in Drosophila melanogaster. Structurally, Debcl is similar to Bax-like proapoptotic Bcl-2 family members. Ectopic expression of Debcl in cultured cells and in transgenic flies causes apoptosis, which is inhibited by coexpression of the baculovirus caspase inhibitor P35, indicating that Debcl is a proapoptotic protein that functions in a caspase-dependent manner. debcl expression correlates with developmental cell death in specific Drosophila tissues. We also show that debcl genetically interacts with diap1 and dark, and that debcl-mediated apoptosis is not affected by gene dosage of rpr, hid, and grim. Biochemically, Debcl can interact with several mammalian and viral prosurvival Bcl-2 family members, but not with the proapoptotic members, suggesting that it may regulate apoptosis by antagonizing prosurvival Bcl-2 proteins. RNA interference studies indicate that Debcl is required for developmental apoptosis in Drosophila embryos. These results suggest that the main components of the mammalian apoptosis machinery are conserved in insects.

Show MeSH
Related in: MedlinePlus