DNA structure modification by the Schlafen protein family

Schlafen 蛋白家族对 DNA 结构的修饰

基本信息

批准号：
9238658
负责人：
MICHAEL DAVID
金额：
$ 18.11万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-07 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9238658
关键词：
ATP phosphohydrolase ATPase Domain Alternative Splicing Antibodies Antineoplastic Agents Antiviral Agents Attention Attenuated Autoimmune Diseases B-DNA Binding Biological Biological Process Biology Caenorhabditis elegans Cancer cell line Cell Line Cells Codon Nucleotides Communicable Diseases DNA DNA Binding DNA Binding Domain DNA Damage DNA Structure Data Derivation procedure Diagnostic Down-Regulation Drosophila genus ERG gene Elements Employee Strikes Family Funding Funding Agency Generations Genes Genetic Goals HIV Hand Human IRF3 gene Immune response Immunologics Infection Infection prevention Innate Immune Response Interferon-alpha Interferons Investigation Kinetics Laboratories Left Length Link Lupus Erythematosus Messenger RNA Modernization Modification N-terminal Nucleic Acid Conformation Nucleic Acids Organism Pathway interactions Pharmacology Physiological Play Poxviridae Process Production Protein Biosynthesis Protein Family Protein Isoforms Proteins Publishing RNA RNA Helicase Research Research Personnel Resistance Resistance development Retroviridae Role Simian virus 40 Structure Surface System Techniques Technology Testing Therapeutic Intervention Time Transfer RNA Vaccine Adjuvant Viral Viral Proteins Virus Virus Diseases Work Yeasts Z-DNA Binding Protein Z-Form DNA adaptive immune response antitumor drug base cancer cell chemotherapeutic agent cytotoxic experimental study genome-wide analysis helicase immunogenic in vivo insight interest member neoplastic cell novel pathogen prevent prospective public health relevance small molecule tool vaccine development

项目摘要

DESCRIPTION (provided by applicant): The innate immune response is not antigen-specific, and is composed of the interferon (IFN) system as well as cell-based anti-pathogen countermeasures that restrict the replication of pathogens. Understanding the innate immune response and the pathways that promote or restrict the kinetics of different steps of infection is essential for devising novel pharmacological strategies for therapeutic intervention during these infectious disease processes, to develop diagnostic tools or to prevent infection. Interferons exert their antiviral effects through the induction of Interferon Stimulated early response Genes (ISGs). An expanding family of such ISGs are the Schlafen (Slfn) genes, which in addition to IFN are also directly induced by pathogen contact with the cells. In our quest to define the biological function of Slfn proteins we discovered that huSlfn11 potently inhibits the production of retroviruses including HIV. Our studies revealed huSlfn11 expression had no effect on the early steps of the infection cycle, but that huSlfn11 protein acts at the very late stages of replication where new viral proteins are synthesized. A striking hallmark of huSlfn11 function is the selective inhibition of the synthesis of virus-encoded proteins, whereas production host cell proteins are apparently unhindered. Most recently, genome-wide screening efforts from two independent laboratories employing dozens of cancer cells lines identified an intriguing correlation between huSlfn11 expression and sensitivity of cells to DDAs. Using cell lines we generated during our previous studies which specifically lack huSlfn11, we find that indeed such cells are resistant to the cytotoxic effects of several classes of DDAs. It is therefore quite reasonable to consider that the development of resistance of tumor cells against DDAs involves the down- regulation of huSlfn11. Finally, our newest studies revealed that huSlfn 5 and 11 have the ability to bind and/or generate Z-DNA. As the structure of the Z-DNA double helix is considerably different from that of B-DNA it supports different interactivity with other molecules, and a few proteins have been identified that selectively bind to Z-DNA but not to B-DNA. It is also notable that unlike B-DNA, Z-DNA is very immunogenic which has not only allowed for the production of highly specific antibodies for research purposes, but has also given rise to anti-Z-DNA antibodies in autoimmune disease such as lupus erythematosus. Although the left-handed Z-DNA and Z-RNA forms were discovered decades ago, their biological role has remained a mystery, largely because little information exists on their formation under physiological conditions. So for several decades not much insight had surfaced on Z-DNA function. In fact, since the end of the 1980s the biology of Z-DNA was not receiving much attention from researchers or funding agencies, and with a few exceptions focused on the role of Z-DNA in genetic instabilities. Our discovery of Z-D/RNA generation by Slfns, a family of interferon-induced antiviral proteins, has the strong potential to place these left-handed nucleic acids into numerous immune response scenarios. Understanding of the biological roles of Z-D/RNA is elementary to exploit their prospective pharmacological possibilities in the development of vaccine adjuvants, anti-viral or anti-neoplastic chemotherapeutics.

描述（由适用提供）：先天免疫响应不是抗原特异性的，并且由干扰素（IFN）系统以及基于细胞的抗病原体对抗来限制病原体的复制。了解先天免疫响应以及促进或限制不同感染步骤动力学的途径对于在这些感染性疾病过程中制定热干预的新型药理学策略至关重要，以开发诊断工具或防止感染。干扰素通过诱导干扰素刺激早期反应基因（ISGS）发挥抗病毒作用。 schlafen（SLFN）基因不断扩展的家族，除了IFN外，该基因也与病原体与细胞接触直接诱导。为了定义SLFN蛋白的生物学功能，我们发现HUSLFN11可能会抑制包括HIV在内的逆转录病毒的产生。我们的研究表明，HUSLFN11表达对感染周期的早期步骤没有影响，但是HUSLFN11蛋白在合成新病毒蛋白的复制阶段起作用。 HUSLFN11功能的惊人标志是对病毒编码蛋白的合成的选择性抑制，而生产宿主细胞蛋白显然受到不受阻碍。最近，使用数十种癌细胞系的两个独立实验室的全基因组筛查工作确定了HUSLFN11表达与细胞对DDA的敏感性之间的有趣相关性。使用我们在先前缺乏HUSLFN11的研究中产生的细胞系，我们发现确实这些细胞对几类DDA的细胞毒性作用具有抗性。因此，考虑到肿瘤细胞对DDA的耐药性的发展涉及HUSLFN11的下调是很合理的。最后，我们的最新研究表明，HUSLFN 5和11具有结合和/或产生Z-DNA的能力。由于Z-DNA双螺旋的结构与B-DNA的结构不同，因此支持与其他分子的不同相互作用。并且已经鉴定出一些蛋白质有选择地结合Z-DNA，但不能与B-DNA结合。同样值得注意的是，与B-DNA不同，Z-DNA具有很高的免疫原性，不仅允许用于研究目的的高度特异性抗体，而且还引起了自身免疫性疾病（例如红斑狼疮）的抗Z-DNA抗体。尽管左撇子Z-DNA和Z-RNA形式是在以前发现的，但它们的生物学作用仍然是神秘的，这主要是因为在物理条件下对它们的形成的信息很少。因此，几十年来，没有太多的见识浮出水面Z-DNA功能。实际上，自1980年代末以来，Z-DNA的生物学并没有得到研究人员或资助机构的太多关注，并且有一些例外的重点是Z-DNA在遗传不稳定性中的作用。我们通过干扰素诱导的抗病毒蛋白SLFNS对Z-D/RNA产生的发现具有很强的潜力，可以将这些左手核酸置于众多的免疫调节情景中。对Z-D/RNA的生物学作用的理解是基本利用其前瞻性药物在疫苗佐剂，抗病毒或抗肿瘤化学治疗药的发展中的前瞻性药物。