• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文学位 >Effect of mutations within the BPV-1 E1 DNA binding domain on E1-E2 interaction and the identification and characterization of E1 interacting peptides.
【24h】

Effect of mutations within the BPV-1 E1 DNA binding domain on E1-E2 interaction and the identification and characterization of E1 interacting peptides.

机译：BPV-1 E1 DNA结合域内的突变对E1-E2相互作用以及E1相互作用肽的鉴定和表征的影响。

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

Interaction of papillomavirus proteins E1 and E2 is critical for replication. In an effort to characterize this important E1-E2 interaction we used a yeast two-hybrid system to determine which domains of each protein were important. This study revealed both an N-terminal E1 (E1N)-E2 and a C-terminal E1 (E1C)-E2 interaction. Upon further mapping of the E1N-E2 interaction, we found that two separate E2 domains, E2TAD (aa 1–194) and E2DBD (aa 326–410) interacted with E1N.; The E1 DNA binding domain (E1DBD), which is found within the E1N domain, contains three hydrophilic regions, HR1, HR2, and HR3. As a means of characterizing the E1N-E2 interaction, we tested HR1, HR2, and HR3 E1DBD mutants for their ability to interact with E2TAD and E2DBD, and to function in a transient replication assay. These data revealed that the HR1 and HR3 regions are important in E2TAD interaction, but a lack of E2TAD interaction did not effect viral replication. Only one E1DBD mutant was found to be defective for E2DBD interaction, and consequently was replication defective. This study also identified a hydrophobic region at the C-terminus of the E1DBD, which appears to play a role in replication.; HPVs have been strongly linked to cervical cancer. One method of treating this disease would be to inhibit viral replication. Recently, a concept for inhibition of replication through peptide interaction has been developed. If a peptide can be identified which interacts with E1 and inhibits one of E1's many functions, it may ultimately affect the ability of E1 to act as the initiator of viral replication, thus inhibiting the ability of the virus to maintain itself within the host cells. In this study we screened a random peptide library and isolated over one hundred E1 interacting peptides. Thirty-two peptides were sequenced and only one peptide was found to be similar to a known E1 interacting protein. All of the peptides assayed interacted with E1N, while no peptide was found to interact solely with E1C. Finally, in an electromobility shift assay no inhibition of E1-DNA or E1-E2 interaction was found in the presence of any of the four peptides tested.
机译：乳头瘤病毒蛋白E1和E2的相互作用对于复制至关重要。为了表征这种重要的E1-E2相互作用，我们使用了酵母双杂交系统来确定每种蛋白质的哪些结构域很重要。这项研究揭示了N端E1（E1N）-E2和C端E1（E1C）-E2相互作用。在进一步映射E1N-E2相互作用后，我们发现两个单独的E2域E2TAD（aa 1–194）和E2DBD（aa 326–410）与E1N相互作用。在E1N域中发现的E1 DNA结合域（E1DBD）包含三个亲水区域HR1，HR2和HR3。作为表征E1N-E2相互作用的一种方法，我们测试了HR1，HR2和HR3 E1DBD突变体与E2TAD和E2DBD相互作用以及在瞬时复制测定中起作用的能力。这些数据表明，HR1和HR3区域在E2TAD相互作用中很重要，但是缺少E2TAD相互作用不会影响病毒复制。发现只有一个E1DBD突变体对E2DBD相互作用有缺陷，因此复制有缺陷。这项研究还确定了E1DBD C端的疏水区，该区似乎在复制中起作用。 HPV与宫颈癌有很强的联系。治疗该疾病的一种方法是抑制病毒复制。最近，已经开发了通过肽相互作用抑制复制的概念。如果可以鉴定出一种与E1相互作用并抑制E1众多功能之一的肽，它可能最终影响E1充当病毒复制起始剂的能力，从而抑制病毒在宿主细胞中维持自身的能力。在这项研究中，我们筛选了一个随机的肽库，并分离了一百多种E1相互作用的肽。对32个肽进行了测序，发现仅一种肽与已知的E1相互作用蛋白相似。所有测定的肽都与E1N相互作用，而没有发现肽仅与E1C相互作用。最后，在电动迁移分析中，在所测试的四种肽中的任何一种的存在下均未发现对E1-DNA或E1-E2相互作用的抑制。

著录项

  • 作者

    Woytek, Kelly Jo.;

  • 作者单位

    Texas A&M University.;

  • 授予单位 Texas A&M University.;
  • 学科 Biology Molecular.; Biology Microbiology.
  • 学位 Ph.D.
  • 年度 2001
  • 页码 113 p.
  • 总页数 113
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;微生物学;
  • 关键词

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