Galili N, Nayak S, Epstein J A, Buck C A
The Wistar Institute, Philadelphia, Pennsylvania 19104, USA.
Dev Dyn. 2000 May;218(1):102-11. doi: 10.1002/(SICI)1097-0177(200005)218:1<102::AID-DVDY9>3.0.CO;2-A.
A yeast 2-hybrid screen was performed to identify possible transcriptional modulators interactive with goosecoid-like (gscl), a transcription factor with suppressive activity, expressed during early brain and gonad development. The screen resulted in the identification of a RING protein known as rnf4 or snrf. Gscl/rnf4 interactions were confirmed by affinity chromatography and by immunoprecipitation. Northern analysis confirmed earlier reports of ubiquitous rnf4 expression in adult tissues. Immunohistochemical analysis of mouse embryos revealed expression primarily in the developing nervous system, with strong expression in the dorsal root ganglia and developing gonads. In contrast to previous reports, both cytoplasmic and nuclear expression of rnf4 was documented. The results reported here confirm and extend earlier reports of rnf4 expression. They suggest for the first time, that in addition to acting as a modulator of transcriptional activity, rnf4 may function, as do other RING proteins, to promote the formation of intracytoplasmic complexes involved in shuttling information between the cytoplasm and the nucleus.
进行了酵母双杂交筛选,以鉴定与类鹅膏蕈氨酸(gscl)相互作用的可能的转录调节因子,gscl是一种具有抑制活性的转录因子,在早期脑和性腺发育过程中表达。筛选结果鉴定出一种名为rnf4或snrf的泛素连接酶蛋白。通过亲和色谱法和免疫沉淀法证实了Gscl/rnf4的相互作用。Northern分析证实了先前关于rnf4在成年组织中普遍表达的报道。对小鼠胚胎的免疫组织化学分析显示,rnf4主要在发育中的神经系统中表达,在背根神经节和发育中的性腺中强烈表达。与先前的报道相反,记录了rnf4的细胞质和细胞核表达。此处报道的结果证实并扩展了先前关于rnf4表达的报道。它们首次表明,除了作为转录活性的调节因子外,rnf4可能像其他泛素连接酶蛋白一样发挥作用,促进参与细胞质和细胞核之间信息穿梭的胞质内复合物的形成。