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CHAPERONE ACTIVITY OF SV40 LARGE T ANTIGEN

SV40 大 T 抗原的伴侣活性

基本信息

批准号：
6497932
负责人：
SHEARA W FEWELL
金额：
$ 4.62万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-01-03 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/6497932
关键词：
DNA replication fungal genetics host organism interaction molecular chaperones neoplastic transformation protein protein interaction protein structure function simian virus 40 stress proteins tumor antigens virion virus assembly virus related neoplasm /cancer virus replication western blottings

项目摘要

The SV40 large tumor antigen (TAg) transforms a variety of cell lines and induces tumors in rodents. In addition, TAg is required to replicate of the SV40 genome, assemble infectious virions and activate the transcription of the SV40 capsid protein genes. While it is clear that these activites require TAg to interact with multiple cellular targets, the mechanism of TAg action and the effect of TAg on these factors is unknown. However, the presence of a functional DnaJ-domain in the amino terminus of TAg suggests that TAg participates with other cellular chaperones to mediate changes in multi-protein complexes involved in these processes. To investigate the chaperone activity of TAg, novel mutations in the J-domain of TAg will be isolated using our recently developed genetic screen in yeast. These mutations will provide insight into the structure-function relationship of the J-domain and will permit further investigation of the role of TAg chaperone activity in SV40 infection and tumorigenesis. Mutations in TAg that compromise its chaperone activity in yeast will be tested in vitro for the ability to stimulate hsp70 and in vivo for the capability to assemble infectious virions, replicate SV40 DNA and transform established cell lines. These experiments will test the "Chaperone Model" for TAg activity by determining whether a functional interaction between TAg and cytosolic hsp7O is required for SV40 infection. The results from these experiments will influence our understanding of virally-induced tumorigenesis and the mechanisms of molecular chaperones in general.

SV40大肿瘤抗原（TAg）在啮齿动物体内转化多种细胞系并诱导肿瘤。此外，还需要TAg来复制SV40基因组，组装感染性病毒粒子，激活SV40衣壳蛋白基因的转录。虽然很明显，这些活动需要TAg与多个细胞靶点相互作用，但TAg的作用机制以及TAg对这些因素的影响尚不清楚。然而，在TAg的氨基端存在一个功能性的dnaj结构域，表明TAg与其他细胞伴侣参与介导参与这些过程的多蛋白复合物的变化。为了研究TAg的伴侣活性，我们将利用我们最近开发的酵母基因筛选技术分离TAg j结构域的新突变。这些突变将有助于深入了解j结构域的结构-功能关系，并允许进一步研究TAg伴侣活性在SV40感染和肿瘤发生中的作用。TAg的突变会损害其在酵母中的伴侣活性，将在体外测试其刺激hsp70的能力，并在体内测试其组装感染性病毒粒子、复制SV40 DNA和转化已建立的细胞系的能力。这些实验将通过确定SV40感染是否需要TAg和细胞质hsp70之间的功能相互作用来测试TAg活性的“伴侣模型”。这些实验的结果将影响我们对病毒诱导的肿瘤发生和分子伴侣的机制的理解。