Zhang Youming, Dean Charlotte, Chessum Lauren, Nguyen Dao, Stewart Michelle, Taylor Martin, Cookson William O, Moffatt Miriam F
Molecular Genetics and Genomics Group, Division of Respiratory Sciences, National Heart and Lung Institute, Imperial College London, Dovehouse Street, London, SW3 6LY, UK,
Mamm Genome. 2014 Dec;25(11-12):573-82. doi: 10.1007/s00335-014-9535-x. Epub 2014 Aug 5.
Previously, human genetic studies have shown association between polymorphisms within the gene encoding plant homeodomain zinc finger protein 11 (PHF11) and asthma-related phenotypes. Initial functional studies have suggested that PHF11 may be involved in the immune response through regulation of T cell activities. In order to study further the gene's functions, we have investigated the mouse Phf11 locus. We have established and characterised a mouse line harbouring a point mutation in the PHD domain of Phf11. Full-length mouse cDNA for Phf11 was obtained by applying rapid amplification of cDNA ends (RACE). All five exons encoding the PHD domain of Phf11 were directly sequenced in 3840 mouse DNA samples from the UK MRC Harwell ENU (N-ethyl-N-nitrosourea)-mutagenised DNA archive. Mice harbouring a valine to alanine substitution, predicted to have a significant functional impact on the PHD zinc finger domain, were re-derived. These Phf11 mutant mice were outcrossed to C3H mice and then backcrossed for ten generations in order to establish a congenic line harbouring the single point mutation in Phf11. Macroscopic examination, haematology and histological examination of lung structure revealed no significant differences between mutant and wild-type mice. After administration of lipopolysaccharide, the level of expression of Il2, NF-kB and Setdb2 were significantly increased in Phf11 mutant homozygous lungs compared to control littermates. Our results provide evidence that Phf11 can operate as a Th1 cell regulator in immune responses. Moreover, our data indicate that these mice may provide a useful model for future studies on Phf11.
此前,人类遗传学研究表明,编码植物同源结构域锌指蛋白11(PHF11)的基因内的多态性与哮喘相关表型之间存在关联。初步功能研究表明,PHF11可能通过调节T细胞活性参与免疫反应。为了进一步研究该基因的功能,我们对小鼠Phf11基因座进行了研究。我们建立并鉴定了一种在Phf11的PHD结构域中存在点突变的小鼠品系。通过应用cDNA末端快速扩增(RACE)获得了Phf11的全长小鼠cDNA。对来自英国医学研究委员会哈韦尔ENU(N-乙基-N-亚硝基脲)诱变DNA文库的3840份小鼠DNA样本中编码Phf11的PHD结构域的所有五个外显子进行了直接测序。携带缬氨酸到丙氨酸取代的小鼠被重新培育,预计该取代会对PHD锌指结构域产生重大功能影响。这些Phf11突变小鼠与C3H小鼠杂交,然后回交十代,以建立一个在Phf11中携带单点突变的近交系。对肺部结构的宏观检查、血液学检查和组织学检查显示,突变小鼠和野生型小鼠之间没有显著差异。给予脂多糖后,与对照同窝小鼠相比,Phf11突变纯合子肺中Il2、NF-kB和Setdb2的表达水平显著升高。我们的结果提供了证据,表明Phf11在免疫反应中可作为Th1细胞调节因子发挥作用。此外,我们的数据表明,这些小鼠可能为未来关于Phf11的研究提供一个有用的模型。