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Updated: Aug 7, 2026
Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death
Published on: October 21, 2012
[The primary study on a novel protein binding to the death domain of the death receptor 4]
Xiao-ling Li1, Yan-xin Liu, Shi-lian Liu
1National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, PUMC and CAMS, Beijing 100005, China.
Objective:
To clone and identify novel proteins binding to the death domain of the death receptor 4 (DR4).
Methods:
The yeast two-hybrid system was used for this study. Automatic sequencing was carried out for DNA sequencing. The sequence homology and the functional domains were analyzed by BLAST and the ScanProsite Tool softwares, respectively. Co-immunoprecipitate method was used to confirm human formyl peptide receptor-like 1 (FPRL1) binding specifically with DR4CD (the cytoplasmic domain of DR4) in HEK293T cells.
Results:
Two positive clones, named as pADB1 and pADB2, were obtained. BLAST searching showed that the homology of the insert sequence of pADB1 with the mRNA of FPRL1 was 97%. The insert of pADB2 shared no homology with any known peptides in GeneBank. Co-immunoprecipitate analysis further confirmed that FPRL1 could bind to DR4CD in vivo specifically.
Conclusions:
FPRL1 may associate with DR4CD in vivo specifically. The functional studies of FPRL1 in signaling pathway mediated by TNF-related apoptosis inducing ligand (TRAIL) are in active progress in our laboratory.
Insights
Researchers identified human formyl peptide receptor-like 1 (FPRL1) as a novel protein that binds to the death receptor 4 (DR4) cytoplasmic domain. This interaction may play a role in apoptosis signaling pathways.
Area of Science:
- Molecular Biology
- Cell Biology
- Immunology
Context:
- Death receptor 4 (DR4) is a key mediator of apoptosis.
- Understanding DR4-interacting proteins is crucial for deciphering cell death pathways.
- Novel protein interactions with DR4 remain to be fully elucidated.
Purpose:
- To clone and identify novel proteins that bind to the death domain of DR4.
- To investigate the specific interaction between identified proteins and the DR4 cytoplasmic domain (DR4CD).
Summary:
- A yeast two-hybrid system identified two positive clones, pADB1 and pADB2.
- Sequence analysis revealed pADB1 insert homology to human formyl peptide receptor-like 1 (FPRL1) mRNA (97%).
- Co-immunoprecipitation confirmed specific in vivo binding of FPRL1 to DR4CD.
Impact:
- Identified FPRL1 as a potential binding partner for DR4.
- Suggests a novel role for FPRL1 in DR4-mediated signaling.
- Provides a foundation for further functional studies of FPRL1 in TNF-related apoptosis-inducing ligand (TRAIL) pathways.
