...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >SUBSTRATE-LENGTH-DEPENDENT ACTIVITIES OF HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 INTEGRASE IN VITRO - DIFFERENTIAL DNA BINDING AFFINITIES ASSOCIATED WITH DIFFERENT LENGTHS OF SUBSTRATES
【24h】

SUBSTRATE-LENGTH-DEPENDENT ACTIVITIES OF HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 INTEGRASE IN VITRO - DIFFERENTIAL DNA BINDING AFFINITIES ASSOCIATED WITH DIFFERENT LENGTHS OF SUBSTRATES

机译：人免疫缺陷病毒1型整合酶在基质不同长度的体外DNA结合亲和力中的底物长度依赖性活动

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Human immunodeficiency virus type 1 integrase (HIV-IN) is an enzyme essential for the integration of viral DNA into the host chromosome, a process that is an attractive target for drug development. In vitro assays have been developed to study both components of the integration process, the 3'-processing and strand transfer reactions, However, major discrepancies between results obtained from in vivo and in vitro events raise concerns as to the biological relevance of activities observed in vitro. These discrepancies include the size of the substrate and the nature of the divalent cation used, In this study, we characterized activities of HIV-IN with oligonucleotide substrates varying in length. Our previous studies indicate that the preferred cation in vitro for 3'-processing is altered from Mn2+ to Mg2+ by increasing the length of the oligonucleotide substrate. This study demonstrates that HIV-IN efficiently catalyzes Mg2+-dependent 3'-processing while repressing the strand transfer reaction. Substrate competition studies indicate that longer substrates preferentially bind Co the viral DNA binding site of the integrase, whereas the shorter substrate has much less specificity. In addition, the shorter substrate requires a higher concentration of Mg2+, indicating that there is an alteration in the metal binding affinity associated with the varying substrates. Our results show that substrate-length-dependent differential activities are due to differences in the divalent metal binding and DNA binding affinities associated with the different substrates. These results suggest that the structure of the viral DNA is an important factor in differentiating the donor and target substrates. Characterization of DNA substrates for in vitro assays may resolve some of the in vitro and in vivo discrepancies and provide further understanding of structure/function relationships between IN and DNA.
机译：人类免疫缺陷病毒1型整合酶（HIV-IN）是将病毒DNA整合到宿主染色体中必不可少的酶，这一过程是药物开发的有吸引力的目标。已经开发了体外测定法来研究整合过程，3'-加工和链转移反应的两个组成部分。但是，体内和体外事件获得的结果之间的主要差异引起了人们对在体内观察到的活性的生物学相关性的担忧。体外。这些差异包括底物的大小和所用二价阳离子的性质。在这项研究中，我们用长度可变的寡核苷酸底物表征了HIV-IN的活性。我们以前的研究表明，通过增加寡核苷酸底物的长度，体外3'处理的优选阳离子从Mn2 +变为Mg2 +。这项研究表明，HIV-IN可有效催化Mg2 +依赖性3'-加工，同时抑制链转移反应。底物竞争研究表明，较长的底物优先结合整合酶的病毒DNA结合位点Co，而较短的底物则特异性低得多。此外，较短的底物需要较高的Mg2 +浓度，表明与不同底物相关的金属结合亲和力发生了变化。我们的结果表明，依赖于底物长度的差异活性是由于与不同底物相关的二价金属结合和DNA结合亲和力的差异。这些结果表明，病毒DNA的结构是区分供体和靶底物的重要因素。用于体外测定的DNA底物的表征可以解决某些体外和体内差异，并提供对IN和DNA之间结构/功能关系的进一步了解。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号