Uptake and Processing of Foreign DNA by Mitochondria, a Model System

描述(申请人提供)：外来DNA进入线粒体的机制在很大程度上是未知的。这项研究的目标是建立一个研究线粒体摄取、复制和转录外源DNA的体外模型。这个基于细胞培养的模型系统将利用一种独特的类病毒制剂，它可以在热带海洋鱼类双色丹自鱼(Stestes Artitus)的肿瘤细胞线粒体中感染和复制。这种传播剂(DVLA)与自然发生的被称为大自鱼性神经纤维瘤病的传染病有关。DVLA与任何已知的病毒或类病毒家族不同。这种制剂DNA基因组的不寻常序列和二级结构可能是这种制剂表面上独特的生活方式的直接结果，并可能为优化线粒体对外来DNA的摄取和处理提供线索。这项研究的目的是检测DVLA DNA进入分离的线粒体，并评估这种DNA在线粒体中的复制和转录。这是一个高风险的研究项目，有可能产生数据，这些数据应该广泛应用于了解线粒体的发病机制，并开发其他脊椎动物的线粒体转基因技术。

DESCRIPTION (provided by applicant): The mechanisms by which foreign DNAs enter mitochondria are largely unknown. The goal of the proposed studies is to develop an in vitro model for study of uptake, replication and transcription of foreign DNA by mitochondria. This cell culture based model system will take advantage of a unique virus-like agent which infects and replicates in the mitochondria of tumor cells in a tropical marine fish, the bicolor damselfish (Stegastes partitus). This transmissible agent (DVLA) is associated with the naturally occurring, infectious disease termed damselfish neurofibromatosis. DVLA does not resemble any known family of virus or viroid. The unusual sequence and secondary structure of the DNA genome of this agent are likely a direct result of the apparently unique life-style of this agent and may provide clues to optimizing uptake and processing of foreign DNA by mitochondria. The aims of the proposed study are to examine the entry of DVLA DNA into isolated mitochondria and evaluate the replication and transcription of this DNA in the mitochondria. This is a high risk research project with the potential to yield data which should be widely applicable to understanding pathogenesis in mitochondria and for development of techniques for mitochondrial transgenesis in other vertebrates.