Related Experiment Video
Updated: Jun 13, 2026
Isolation of Group 2 Innate Lymphoid Cells from Mouse Nasal Mucosa to Detect the Expression of CD226
Published on: May 10, 2022
Molecular characterization of the canine cytokine TWEAK (CD255) and its receptor, Fn14 (CD266)
Jia-Xin Zhang1, Ming Sang, Wei Zhao
1Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210097, PR China.
Abstract:
TWEAK is a member of the tumor necrosis factor superfamily. The interaction of TWEAK with its receptor Fn14 regulates multiple cellular responses, including stimulation of proliferation, migration, apoptosis, angiogenesis, and induction of proinflammatory cytokines. This paper reports for the first time the molecular cloning of dog TWEAK and Fn14. The open reading frame (ORF) of dog TWEAK is 750bp, its genomic DNA consists of seven exons and six introns and is approximately 10kb in size by computer-assisted analysis. Sequence similarity at the amino acid level between dog TWEAK and human or mouse was 95 and 92%, respectively. The ORF of dog Fn14 contains 390bp. Sequence similarity at the amino acid level between dog Fn14 and human, or mouse, or frog was 95, 93 and 64%, respectively. Real-time quantitative PCR analysis revealed that both TWEAK and Fn14 are constitutively expressed in various tissues in dog. Furthermore, we verified dTWEAK interacted with dFn14 by yeast two-hybrid assay. Our results suggest that the TWEAK-Fn14 pathway is evolutionarily highly conserved. It will be helpful for investigation on the biological role of the TWEAK/Fn14 system in this important animal model. Furthermore, it provides insight into the molecular evolution of the emerging TWEAK and Fn14 families.
Insights
Researchers cloned dog TWEAK and Fn14, revealing high similarity to human and mouse proteins. This TWEAK-Fn14 pathway is conserved in dogs, offering a model for studying its biological roles and evolution.
Area of Science:
- Molecular Biology
- Immunology
- Comparative Genomics
Background:
- TWEAK (TNF-like weak inducer of apoptosis) is a cytokine belonging to the tumor necrosis factor superfamily.
- Its receptor, Fn14, mediates cellular responses like proliferation, migration, apoptosis, angiogenesis, and cytokine induction.
- Understanding the TWEAK-Fn14 system in diverse species is crucial for biological and evolutionary insights.
Purpose of the Study:
- To molecularly clone and characterize dog TWEAK and Fn14.
- To investigate the expression patterns of dog TWEAK and Fn14 in various tissues.
- To confirm the interaction between dog TWEAK and dog Fn14.
Main Methods:
- Molecular cloning of dog TWEAK and Fn14 genes.
- Bioinformatic analysis of gene sequences and structures.
- Real-time quantitative PCR for gene expression analysis.
- Yeast two-hybrid assay to verify protein-protein interaction.
Main Results:
- The open reading frame (ORF) of dog TWEAK is 750bp, with genomic DNA spanning approximately 10kb across seven exons and six introns.
- Dog TWEAK shares 95% and 92% amino acid similarity with human and mouse TWEAK, respectively.
- The ORF of dog Fn14 is 390bp, showing 95%, 93%, and 64% amino acid similarity to human, mouse, and frog Fn14, respectively.
- Both dog TWEAK and Fn14 are constitutively expressed in various dog tissues.
- The interaction between dog TWEAK (dTWEAK) and dog Fn14 (dFn14) was confirmed via yeast two-hybrid assay.
Conclusions:
- The TWEAK-Fn14 signaling pathway is highly conserved across species, including dogs.
- The molecular cloning and characterization of dog TWEAK and Fn14 provide valuable tools for studying this system in a significant animal model.
- These findings offer insights into the molecular evolution of the TWEAK and Fn14 families.
