TISSUE SPECIFIC TUMOR INDUCTION BY POLYOMAVIRUS

多瘤病毒诱导组织特异性肿瘤

基本信息

批准号：
2871823
负责人：
JAN CERNY
金额：
$ 10.71万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-02-01 至 2001-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2871823
关键词：
DNA binding protein DNA footprinting Polyomavirus affinity chromatography gel mobility shift assay gene mutation genetic enhancer element genetic regulatory element laboratory mouse nucleic acid sequence oligonucleotides oncogenic virus thymus neoplasms viral carcinogenesis virus genetics virus infection mechanism virus related neoplasm /cancer

项目摘要

Polyomavirus normally establishes a silent, persistent infection in its natural host, the mouse, but under experimental conditions can cause widespread lytic infection and induce tumors in up to a dozen different tissues, including a high frequency of tumors in thymic tissue, salivary glands and mammary glands. Although the virus has been studied extensively in cell culture, little is known about its interaction with host cells in the animal. The experiments in this proposal are designed to explore the virus-cell interactions that result in tissue specificity of tumor formation by polyomavirus. The two specific aims in this proposal are based on the hypothesis that viral regulatory sequences interacting with cellular factors are responsible for tissue specific tumor induction. The first goal is to define the viral genetic determinants responsible for tissue specific tumorigenesis by evaluating the tumor profile of viruses containing mutations in potential regulatory sequences. These regulatory regions include a 4Obp duplication of sequences on the early side of the origin of replication that we have previously shown to be necessary for the induction of thymic epitheliomas, and the polyoma enhancer sequences that are important for virus regulation in cell culture systems. Our second goal is to use these regulatory sequences to identify and characterize the cellular factors that bind to them. The DNA binding proteins will be identified by gel shift and footprinting experiments and further characterized by UV crosslinking experiments and affinity chromatography. Polyomavirus gene expression and replication, as well as the oncogenic pathways triggered by the virus, utilize cellular factors that are normally involved in signal transduction and cell growth control, and that are altered in many cancers of non-viral etiology. Thus, understanding tumor induction by polyomavirus at the molecular level and identifying cis-acting sequences and cellular factors involved in the tissue specificity of tumor formation will provide new information relevant to the fields of cell growth control and cancer.

多瘤病毒通常在其中建立一个沉默的，持续的感染天然寄主，小鼠，但在实验条件下可能导致广泛的裂解感染并诱导多达十二种不同的肿瘤组织，包括胸腺组织中肿瘤的高频，唾液腺体和乳腺。尽管已广泛研究了该病毒在细胞培养中，它与宿主细胞中的相互作用知之甚少动物。本提案中的实验旨在探索导致肿瘤组织特异性的病毒 - 细胞相互作用多瘤病毒形成。该提案中的两个具体目标是基于以下假设。与细胞因子相互作用的病毒调节序列是负责组织特异性肿瘤诱导。第一个目标是定义负责组织特异性的病毒遗传决定因素肿瘤发生通过评估含有含有病毒的肿瘤特征潜在调节序列中的突变。这些监管区域在起源的早期一侧包括4OBP的序列重复我们以前证明的复制是催化性上皮瘤的诱导和多瘤增强子序列对于细胞培养系统中的病毒调节很重要。我们的第二个目标是使用这些调节序列来识别和表征与它们结合的细胞因子。 DNA结合蛋白将是通过凝胶移位和足迹实验确定以紫外线交联实验和亲和色谱法为特征。多瘤病毒基因表达和复制，以及致癌病毒触发的途径，利用了细胞因子通常参与信号转导和细胞生长控制，并且在许多非病毒病因的癌症中都改变了。因此，理解多瘤病毒在分子水平上诱导肿瘤并鉴定涉及组织中的顺式作用序列和细胞因子肿瘤形成的特异性将提供与细胞生长控制和癌症领域。