Azzalin Alberto, Del Vecchio Igor, Ferretti Luca, Comincini Sergio
Dipartimento di Genetica e Microbiologia, Università di Pavia, via Ferrata 1, 27100 Pavia, Italy.
Anticancer Res. 2006 Nov-Dec;26(6B):4539-47.
Doppel (Dpl) is a homologue of the prion protein (PrPC). In contrast to PrP(C), Dpl is dispensable for prion disease, but appears to have an essential function in male spermatogenesis. Recently, Dpl has been found to be aberrantly expressed in astrocytic and leukaemic tumor specimens, showing a peculiar cytosolic cellular localization. The aim of this study was to clarify some of the putative Dpl interacting proteins.
A yeast two hybrid system was employed and the results were verified by co-immunoprecipitation using transfected cells.
Several potential Dpl-binding candidates were identified and, among them, the receptor for activated C-kinase (RACK1) protein was further investigated. RACK1 deletion mutants showed that some of its WD containing domains were directly involved in the binding with Dpl. Our data showed that Dpl interacts with RACK1 by means of its structured globular carboxyl-terminal region.
This new Dpl interacting partner might suggest functional hypotheses about the role of this protein in an astrocytoma context where Dpl was found ectopically expressed.
多普蛋白(Dpl)是朊病毒蛋白(PrPC)的同源物。与PrP(C)不同,Dpl对于朊病毒疾病并非必需,但似乎在雄性精子发生中具有重要功能。最近,已发现Dpl在星形细胞和白血病肿瘤标本中异常表达,呈现出特殊的胞质细胞定位。本研究的目的是阐明一些假定的与Dpl相互作用的蛋白质。
采用酵母双杂交系统,并使用转染细胞通过免疫共沉淀验证结果。
鉴定出几个潜在的与Dpl结合的候选物,其中,对活化C激酶受体(RACK1)蛋白进行了进一步研究。RACK1缺失突变体表明其一些含WD的结构域直接参与与Dpl的结合。我们的数据表明,Dpl通过其结构化的球状羧基末端区域与RACK1相互作用。
这个新的与Dpl相互作用的伙伴可能提示关于该蛋白在发现Dpl异位表达的星形细胞瘤背景中的作用的功能假说。