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Molecular Genetics of Parvoviral DNA Replication

细小病毒 DNA 复制的分子遗传学

基本信息

批准号：
7846562
负责人：
Peter J. Tattersall
金额：
$ 0.91万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-05 至 2009-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7846562
关键词：
Alien Binding Binding Sites Biochemical Capsid Cell Culture System Cell Culture Techniques Cell Nucleus Cells Childhood Communicable Diseases DNA DNA Damage DNA biosynthesis Ear Elements Frequencies Genes Genetic Genome Goals Host Defense Human Human Virus Infection Knowledge Learning Left Malignant Neoplasms Maps Mice Minute Virus Modeling Molecular Molecular Genetics Molecular Motors Motor Mus Mutant Strains Mice Natural History Nature Nucleoproteins Nucleosomes Organism PTPN11 gene Parvovirus Population Process Proteins Replication Initiation Replication Origin Research Role Structure Subgroup System Terminal Repeat Sequences Transgenes Viral Viral Genome Virus chromatin immunoprecipitation in vivo melting mouse genome mutant pathogen positional cloning programs public health relevance response telomere vector vector vaccine viral DNA

项目摘要

DESCRIPTION (provided by applicant): Parvoviruses are unique among all known organisms in having DNA genomes which are both single-stranded and linear. Both 5' and 3' ends of the genome are palindromic and can fold into hairpin structures that give rise to the two viral replication origins in replicative-form (RF) DNA. In the homotelomeric parvoviruses these hairpins are part of a terminal repeat, whereas for the heterotelomeric subgroup, the termini are typically unrelated to one another in either sequence or structure. The long term goal of this research program is to understand, at the molecular level, how heterotelomeric parvoviruses inveigle their host cell into replicating and packaging such apparently alien molecules. The major model we will continue to explore is the autonomous parvovirus Minute Virus of Mice (MVM), a genetically tractable virus that grows in cell culture. We will use what we learn from MVM to explore the function of the initiator protein of the newly-discovered pathogenic human bocavirus HBoV, for which there is currently no cell culture system. We propose to build on our previous research on MVM DNA replication initiation to study the structure and function of the terminal hairpins, extending this to the precise sequence requirements for NS1 binding, melting and nicking at MVM OriL. We will explore the functional significance of NS1 binding sites embedded in the MVM genome, and ask whether a similar situation pertains for HBoV. We will initiate studies on how the virus activates cellular DNA damage responses and whether the unique pseudochromatin that the virus elaborates is part of its strategy to evade, or employ, these elements of innate host defense. Genetic and biochemical approaches will be pursued to identify the molecular motor that drives genome packaging, and to explore the function(s) of the teminal hairpins in viral DNA replication and packaging. PUBLIC HEALTH RELEVANCE: This research program aims to develop an understanding of the mechanisms underlying parvoviral DNA replication and packaging, and further our knowledge of parvoviruses as pathogens. These are also necessary prerequisites for the use of these viruses as vaccine vectors against infectious diseases and cancer. The newly discovered human parvovirus, HBoV, appears to be one of the most prevalent infections of childhood, and understanding its natural history and mode of replication will become of increasing importance as we discover more about its interaction with the human population. In the present renewal application, therefore, we propose to parlay what we know about parvoviral genome structure and DNA replication from the murine system into a study of similar processes that regulate replication of the human virus.

描述(申请人提供)：细小病毒在所有已知的生物体中是独一无二的，因为它的DNA基因组既是单链的，也是线性的。基因组的5‘和3’端都是回文的，可以折叠成发夹结构，从而在复制形式(RF)DNA中产生两个病毒复制起点。在同体细小病毒中，这些发夹是末端重复的一部分，而对于异体亚群，末端通常在序列或结构上彼此无关。这项研究计划的长期目标是在分子水平上了解异体细小病毒如何诱使宿主细胞复制和包装这种表面上是外来的分子。我们将继续探索的主要模型是小鼠自主细小病毒微小病毒(MVM)，这是一种在细胞培养中生长的遗传易受感染的病毒。我们将利用从MVM学到的东西来探索新发现的致病性人类博卡病毒HBoV的启动子蛋白的功能，目前还没有针对这种病毒的细胞培养系统。我们建议在我们以前对MVM DNA复制起始的研究的基础上，研究末端发夹的结构和功能，将其扩展到MVM OriL上NS1结合、熔化和缺口的精确序列要求。我们将探索嵌入在MVM基因组中的NS1结合位点的功能意义，并询问类似的情况是否适用于HBoV。我们将开始研究病毒如何激活细胞DNA损伤反应，以及病毒所阐述的独特的假染色质是否是其逃避或利用这些固有宿主防御元素的策略的一部分。将采用遗传和生化方法来确定驱动基因组包装的分子马达，并探索末端发夹在病毒DNA复制和包装中的功能(S)。公共卫生相关性：该研究计划旨在了解细小病毒DNA复制和包装的潜在机制，并进一步了解细小病毒作为病原体的知识。这些也是将这些病毒用作对抗传染病和癌症的疫苗载体的必要先决条件。新发现的人类细小病毒HBoV似乎是儿童最常见的感染之一，随着我们发现更多关于它与人类人口的相互作用，了解它的自然历史和复制模式将变得越来越重要。因此，在目前的更新应用中，我们建议将我们所知的微小病毒基因组结构和DNA复制从小鼠系统转移到调节人类病毒复制的类似过程的研究中。

项目成果

期刊论文数量（38）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structure of a packaging-defective mutant of minute virus of mice indicates that the genome is packaged via a pore at a 5-fold axis.

小鼠微小病毒包装缺陷突变体的结构表明，基因组是通过5倍轴上的孔进行包装的。

DOI：
10.1128/jvi.02598-10
发表时间：
2011
期刊：
Journal of virology
影响因子：
5.4
作者：
Plevka,Pavel;Hafenstein,Susan;Li,Lei;D'AbrgamoJr,Anthony;Cotmore,SusanF;Rossmann,MichaelG;Tattersall,Peter
通讯作者：
Tattersall,Peter

Expression of functional parvoviral NS1 from recombinant vaccinia virus: effects of mutations in the nucleotide-binding motif.