The Caenorhabditis elegans CED-9 protein does not directly inhibit the caspase CED-3, in vitro nor in yeast

Jabbour, Anissa; Ho, Po Ki; Puryer, Michelle A; Ashley, David M.; Ekert, Paul G.; Hawkins, Christine J.

doi:10.1038/sj.cdd.4401501

Cited by 12 publications

(20 citation statements)

References 29 publications

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“…As we reported previously (Jabbour et al, 2004), yeast survived expression of either CED-3 or CED-4 alone (Fig. 1A, lanes 2 and 3).…”

Section: Resultssupporting

confidence: 87%

“…Here, we capitalised on the previously noted ability of active caspases to kill the yeast Saccharomyces cerevisiae (Hawkins et al, 2001;Hawkins et al, 1999;Hawkins et al, 2000a;Hawkins et al, 2000b;Jabbour et al, 2002;Jabbour et al, 2004;Kang et al, 1999), to reconstitute the core nematode apoptotic pathway and to explore the ability of Bcl-2 to interact with it. Caspase toxicity in yeast depends both on the enzyme adopting an active conformation, as well as its substrate specificity.…”

Section: Resultsmentioning

confidence: 99%

“…1-251 and pADH-(TRP1)-FLAG-CED-9 1-251 have been previously described Jabbour et al, 2004). pCUP1-(TRP1) was generated by subcloning the CUP1 promotor, polylinker and actin terminator ApaI-NotI fragment from pCUP1-(LEU2) into pRS314 (Sikorski and Hieter, 1989) cut with ApaI and NotI, to give pCUP1-(TRP1).…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…was amplified using oligonucleotides 6 and 7 from p416-MET25-GFP S65T (Jabbour et al, 2004) and cut with EcoRI. The digested PCR product was then ligated into both pADH-(TRP1)-CED-9 and pADH-(TRP1)-CED-9…”

Section: S65tmentioning

confidence: 99%

“…Bacterial expression constructs pET23a-p35 has been previously reported (Jabbour et al, 2004). EGL-1 was amplified using oligonucleotides 21 and 22.…”

Section: S65tmentioning

confidence: 99%

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Human Bcl-2 cannot directly inhibit the Caenorhabditis elegans Apaf-1 homologue CED-4, but can interact with EGL-1

Jabbour

Puryer

et al. 2006

Journal of Cell Science

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Although the anti-apoptotic activity of Bcl-2 has been extensively studied, its mode of action is still incompletely understood. In the nematode Caenorhabditis elegans, 131 of 1090 somatic cells undergo programmed cell death during development. Transgenic expression of human Bcl-2 reduced cell death during nematode development, and partially complemented mutation of ced-9, indicating that Bcl-2 can functionally interact with the nematode cell death machinery. Identification of the nematode target(s) of Bcl-2 inhibition would help clarify the mechanism by which Bcl-2 suppresses apoptosis in mammalian cells. Exploiting yeast-based systems and biochemical assays, we analysed the ability of Bcl-2 to interact with and regulate the activity of nematode apoptosis proteins. Unlike CED-9, Bcl-2 could not directly associate with the caspase-activating adaptor protein CED-4, nor could it inhibit CED-4-dependent yeast death. By contrast, Bcl-2 could bind the C. elegans pro-apoptotic BH3-only Bcl-2 family member EGL-1. These data prompt us to hypothesise that Bcl-2 might suppress nematode cell death by preventing EGL-1 from antagonising CED-9, rather than by inhibiting CED-4.

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“…As we reported previously (Jabbour et al, 2004), yeast survived expression of either CED-3 or CED-4 alone (Fig. 1A, lanes 2 and 3).…”

Section: Resultssupporting

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Section: Plasmid Constructionmentioning

confidence: 99%

Section: S65tmentioning

confidence: 99%

“…Bacterial expression constructs pET23a-p35 has been previously reported (Jabbour et al, 2004). EGL-1 was amplified using oligonucleotides 21 and 22.…”

Section: S65tmentioning

confidence: 99%

See 3 more Smart Citations

Human Bcl-2 cannot directly inhibit the Caenorhabditis elegans Apaf-1 homologue CED-4, but can interact with EGL-1

Jabbour

Puryer

et al. 2006

Journal of Cell Science

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Current awareness on yeast

2005

Yeast

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In order to keep subscribers up‐to‐date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly‐published material on yeasts. Each bibliography is divided into 10 sections. 1 Books, Reviews & Symposia; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (5 weeks journals ‐ search completed 12th. Jan. 2005)

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Human, insect and nematode caspases kill Saccharomyces cerevisiae independently of YCA1 and Aif1p

Puryer

Hawkins

2006

Apoptosis

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This study characterised the impact of active metazoan apoptotic proteases (caspases) on Saccharomyces cerevisiae viability. Expression of active caspase-3 or caspase-8 in yeast ruptured plasma and nuclear membranes and dramatically impaired clonogenic survival, but did not damage DNA. Deletion of the proposed yeast apoptosis regulators YCA1 or Aif1p did not affect the ability of human, insect or nematode caspases to kill yeast. These data indicate that expression of active metazoan caspases causes irreversible damage to yeast membranes and organelles, in a manner independent of YCA1 and Aif1p.

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The Caenorhabditis elegans CED-9 protein does not directly inhibit the caspase CED-3, in vitro nor in yeast

Cited by 12 publications

References 29 publications

Human Bcl-2 cannot directly inhibit the Caenorhabditis elegans Apaf-1 homologue CED-4, but can interact with EGL-1

Human Bcl-2 cannot directly inhibit the Caenorhabditis elegans Apaf-1 homologue CED-4, but can interact with EGL-1

Current awareness on yeast

Human, insect and nematode caspases kill Saccharomyces cerevisiae independently of YCA1 and Aif1p

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