Zhang Jia-Xin, Sang Ming, Zhao Wei, Ai Hong-Xin, Shui Yan, Li Jian-Feng, Song Ren, Zhang Shuang-Quan
Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210097, PR China.
Vet Immunol Immunopathol. 2010 Sep 15;137(1-2):172-5. doi: 10.1016/j.vetimm.2010.04.004. Epub 2010 Apr 14.
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.
肿瘤坏死因子样弱凋亡诱导因子（TWEAK）是肿瘤坏死因子超家族的成员。TWEAK与其受体Fn14的相互作用可调节多种细胞反应，包括刺激增殖、迁移、凋亡、血管生成以及诱导促炎细胞因子。本文首次报道了犬TWEAK和Fn14的分子克隆。犬TWEAK的开放阅读框（ORF）为750bp，通过计算机辅助分析，其基因组DNA由7个外显子和6个内含子组成，大小约为10kb。犬TWEAK与人或小鼠在氨基酸水平的序列相似性分别为95%和92%。犬Fn14的ORF包含390bp。犬Fn14与人、小鼠或青蛙在氨基酸水平的序列相似性分别为95%、93%和64%。实时定量PCR分析显示，TWEAK和Fn14在犬的各种组织中均组成性表达。此外，我们通过酵母双杂交试验验证了犬TWEAK与犬Fn14相互作用。我们的结果表明，TWEAK-Fn14途径在进化上高度保守。这将有助于在这个重要的动物模型中研究TWEAK/Fn14系统的生物学作用。此外，它还为新兴的TWEAK和Fn14家族的分子进化提供了见解。
