Lindskog Cecilia, Kuhlwilm Martin, Davierwala Armaity, Fu Ning, Hegde Geeta, Uhlén Mathias, Navani Sanjay, Pääbo Svante, Pontén Fredrik
Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University , Dag Hammarskjölds väg 20, SE-751 85 Uppsala, Sweden.
J Proteome Res. 2014 Aug 1;13(8):3596-606. doi: 10.1021/pr500045f. Epub 2014 Jul 15.
RUNX2, a gene involved in skeletal development, has previously been shown to be potentially affected by positive selection during recent human evolution. Here we have used antibody-based proteomics to characterize potential differences in expression patterns of RUNX2 interacting partners during primate evolution. Tissue microarrays consisting of a large set of normal tissues from human and macaque were used for protein profiling of 50 RUNX2 partners with immunohistochemistry. Eleven proteins (AR, CREBBP, EP300, FGF2, HDAC3, JUN, PRKD3, RUNX1, SATB2, TCF3, and YAP1) showed differences in expression between humans and macaques. These proteins were further profiled in tissues from chimpanzee, gorilla, and orangutan, and the corresponding genes were analyzed with regard to genomic features. Moreover, protein expression data were compared with previously obtained RNA sequencing data from six different organs. One gene (TCF3) showed significant expression differences between human and macaque at both the protein and RNA level, with higher expression in a subset of germ cells in human testis compared with macaque. In conclusion, normal tissues from macaque and human showed differences in expression of some RUNX2 partners that could be mapped to various defined cell types. The applied strategy appears advantageous to characterize the consequences of altered genes selected during evolution.
RUNX2是一个参与骨骼发育的基因，此前已被证明在近代人类进化过程中可能受到正选择的影响。在此，我们利用基于抗体的蛋白质组学技术来表征灵长类动物进化过程中RUNX2相互作用伙伴表达模式的潜在差异。由大量人类和猕猴正常组织组成的组织微阵列用于通过免疫组织化学对50个RUNX2伙伴进行蛋白质谱分析。11种蛋白质（AR、CREBBP、EP300、FGF2、HDAC3、JUN、PRKD3、RUNX1、SATB2、TCF3和YAP1）在人类和猕猴之间表现出表达差异。这些蛋白质在黑猩猩、大猩猩和猩猩的组织中进一步进行了分析，并且对相应基因的基因组特征进行了分析。此外，将蛋白质表达数据与先前从六个不同器官获得的RNA测序数据进行了比较。一个基因（TCF3）在蛋白质和RNA水平上在人类和猕猴之间均表现出显著的表达差异，与猕猴相比，在人类睾丸的一部分生殖细胞中表达更高。总之，猕猴和人类的正常组织在一些RUNX2伙伴的表达上存在差异，这些差异可以映射到各种定义的细胞类型。所应用的策略似乎有利于表征进化过程中选择的基因改变的后果。
