Protein-protein interactions between Epstein-Barr virus nuclear antigen-LP and cellular gene products: Binding of 70-kilodalton heat shock proteins
Protein-protein interactions between Epstein-Barr virus nuclear antigen-LP and cellular gene products: Binding of 70-kilodalton heat shock proteins
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DOI：
10.1006/viro.1996.0289
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发表时间：
1996-06-01
期刊：
影响因子：
3.7
通讯作者：
Rowe, DT
中科院分区：
文献类型：
--
作者：
Kitay, MK;Rowe, DT
由Epstein-Barr核抗原信息的前导外显子中的开放阅读框编码的EBNA-LP蛋白对于淋巴细胞的有效永生化是必需的。使用来自[S-35]甲硫氨酸标记的细胞裂解物和细菌表达的麦芽糖结合蛋白EBNA-LP融合物的蛋白质的亲和沉淀进行蛋白质-蛋白质相互作用研究。检测到细胞68/72-kDa的双联体蛋白。该带型与谷胱甘肽-S-转移酶和Sp1（一种基础转录因子）融合的亲和沉淀中获得的带型相同。对于EBNA-LP和Sp1，特异性相互作用的细胞蛋白质已被鉴定为热休克蛋白（HSP）72/73。HSP 72/73与EBNA-LP缺失突变体的亲和沉淀将EBNA-LP上的相互作用结构域映射到永生化所需的外显子Y2。免疫沉淀EBNA-LP从EBV阳性淋巴母细胞系共沉淀HSP 72/73蛋白，表明在体内以及在体外发生的相互作用。热休克蛋白与越来越多的参与基因表达和增殖控制的核蛋白的关联现在包括Sp1和EBNA-LP，并表明分子伴侣在这些过程中起着核心作用。(C)出版社：Academic Press，Inc.
The EBNA-LP protein encoded by the open reading frame in the leader exons of the Epstein-Barr nuclear antigen messages is essential for efficient immortalization of a lymphocytes. Protein-protein interaction studies using affinity precipitation of proteins from [S-35]methionine-labeled cell lysates and bacterially expressed maltose binding protein EBNA-LP fusions were performed. A cellular 68/72-kDa doublet protein was detected. This banding pattern was shown to be identical to that obtained in affinity precipitations with fusions of glutathione-S-transferase and Sp1 (a basal transcription factor). For both EBNA-LP and Sp1 the specific interacting cellular proteins have been identified as heat shock proteins (HSP) 72/73. Affinity precipitation of HSP 72/73 with deletion mutants of EBNA-LP maps the interaction domain on EBNA-LP to exon Y2 which is required for immortalization. Immunoprecipitation of EBNA-LP from EBV-positive lymphoblastoid cell lines coprecipitated the HSP 72/73 proteins, indicating that the interaction occurs in vivo as well as in vitro. The association of HSPs with a widening range of nuclear proteins involved in gene expression and proliferation control now includes Sp1 and EBNA-LP and suggests that there is a central role for molecular chaperones in these processes. (C) 1996 Academic Press, Inc.