Related Experiment Videos
Protection against Fas/APO-1- and tumor necrosis factor-mediated cell death by a novel protein, sentrin
1Division of Molecular Medicine, The University of Texas-Houston Health Science Center 77030, USA.
Abstract:
Fas/APO-1 and TNF receptor 1 share a common signaling motif in their cytoplasmic tail called the "death domain." Using the death domain as bait in the yeast two-hybrid system, several death domain-containing proteins that participate in cell death signaling have been identified. Here we report the isolation of a novel protein, sentrin, which interacts with Fas/APO-1 and TNF receptor 1 but not with FADD/MORT1 or CD40. Two-hybrid interaction assays reveal that sentrin associates only with the signal-competent forms of Fas/APO-1 or TNF receptor 1 death domains. Sentrin is a novel protein of 101 amino acids with homology to ubiquitin, Nedd8, and a Saccharomyces cerevisiae protein, Smt3. When overexpressed, sentrin provides protection against both anti-Fas/APO-1 and TNF-induced cell death.
Insights
A novel protein, sentrin, interacts with Fas/APO-1 and TNF receptor 1 death domains. Overexpression of sentrin protects cells from Fas/APO-1 and TNF-induced cell death, suggesting a role in regulating apoptosis.
Area of Science:
- Molecular Biology
- Cell Biology
- Immunology
Background:
- Fas/APO-1 and TNF receptor 1 are key mediators of programmed cell death.
- These receptors share a cytoplasmic death domain crucial for signal transduction.
- Previous studies identified proteins interacting with death domains using yeast two-hybrid systems.
Purpose of the Study:
- To identify novel proteins that interact with the death domains of Fas/APO-1 and TNF receptor 1.
- To characterize the interaction of the novel protein, sentrin, with these receptors.
- To investigate the functional role of sentrin in cell death signaling.
Main Methods:
- Yeast two-hybrid system was employed using the death domain as bait.
- Interaction assays were performed to confirm binding specificity.
- Protein homology searches were conducted.
- Overexpression studies in cell culture were used to assess functional effects.
Main Results:
- A novel protein, named sentrin, was identified through yeast two-hybrid screening.
- Sentrin specifically interacts with the death domains of Fas/APO-1 and TNF receptor 1.
- Sentrin demonstrated homology to ubiquitin, Nedd8, and Smt3.
- Overexpression of sentrin conferred protection against Fas/APO-1 and TNF-induced cell death.
Conclusions:
- Sentrin is a novel death domain-interacting protein.
- Sentrin plays a protective role in Fas/APO-1 and TNF-mediated apoptosis.
- Sentrin's homology to ubiquitin-like proteins suggests a potential role in post-translational modification pathways regulating cell death.