Interaction between gene family Pax7 and gene coding protein Ubc9
摘要: 目的 了解Pax7的重要生物学功能，筛选并验证与其相互作用的Ubc9基因编码蛋白，并明确Pax7与小泛素相关修饰蛋白（SUMO）的关系，为进一步研究Pax7的作用机制奠定基础。方法 酵母双杂交系统筛选出与Pax7相互作用的基因Ubc9；GST-pull down方法验证Pax7与Ubc9在体外相互作用；Western blot方法检测Pax7的SUMO化状态；免疫共沉淀方法验证Pax7与SUMO的关系。结果 Pax7和Ubc9在酵母中存在相互作用；Pax7和Ubc9在体外可特异性结合；在Ubc9存在的条件下，Pax7呈现SUMO化状态，且可与SUMO相互作用；Pax7在鸡胚胎中也呈现SUMO化状态。结论 Pax7可以与Ubc9相互作用，且这种作用与SUMO有关。
Abstract: Objective To explore biological function of gene family Pax7,its interaction with gene coding protein Ubc9,and the relationship between Pax7 and small ubiquitin-related modifier(SUMO).Methods Yeast two-hybrid was applied to screen out Ubc9 interacted with Pax7.The interaction between Pax7 and Ubc9 in vitro was confirmed with glutathione-S-transferase(GST) pull-down.Western blot was used to detect the SUMOylation of Pax7.The interaction between Pax7 and SUMO was demonstrated with co-immunoprecipitation(Co-IP).Results There was an interaction between Pax7 and Ubc9 in yeast.Pax7 can bind to Ubc9 specially in vitro.Pax7 could be SUMOylated in the presence of Ubc9 and it also could interact with SUMO.Conclusion The results demonstrate that Pax7 interacts with Ubc9,and the interaction leads to the SUMOylation of Pax7.
- Pax7 /
- Ubc9 /
 Paixo-Cǒrtes VR,Salzano FM,Bortolini MC.Evolutionary history of chordate PAX genes:dynamics of change in a complex gene family[J].PloS One,2013,8(9):e73560.  Maczkowiak F,Matos S,Wang Ee,et al.The Pax3 and Pax7 paralogs cooperate in neural and neural crest patterning using distinct molecular mechanisms,in Xenopus laevis embryos[J].Developmental Biology,2010,340(2):381-396.  von Maltzahn J,Jones AE,Parks RJ,et al.Pax7 is critical for the normal function of satellite cells in adult skeletal muscle[J].Proceedings of the National Academy of Sciences of the United States of America,2013,110(41):16474-16479.  罗星,张峪涵,于娜,等.神经突起因子酵母双杂交诱饵质粒构建和转化[J].中国公共卫生,2006,22(4):444-445.  Vadasz S,Marquez,Tulloch M,et al.Pax7 is regulated by cMyb during early neural crest development through a novel enhancer[J].Development(Cambridge,England),2013,140(17):3691-3702.  Murdoch B,DelConte C,GarcaCastro MI.Pax7 lineage contributions to the mammalian neural crest[J].PLoS One,2012,7:e41089.  Betters E,Liu Y,Kjaeldgaard A,et al.Analysis of early human neural crest development[J].Developmental Biology,2010,344(2):578-592.  Voelkel-Meiman K,Taylor LF,Mukherjee P,et al.SUMO localizes to the central element of synaptonemal complex and is required for the full synapsis of meiotic chromosomes in budding yeast[J].PLoS Genetics,2013,9(10):e1003837.  Capili AD,Lima CD.Structure and analysis of a complex between SUMO and Ubc9 illustrates features of a conserved E2-Ubl interaction[J].Journal of Molecular Biology,2007,369(3):608-618.  Klug H,Xaver M,Chaugule VK,et al.Ubc9 sumoylation controls SUMO chain formation and meiotic synapsis in Saccharomyces cerevisiae[J].Molecular Cell,2013,50(5):625-636.
- 文章访问数: 633
- HTML全文浏览量: 122
- PDF下载量: 38
- 被引次数: 0