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Nanoscale Characterization of Interaction of APOBEC3G with RNA

机译：纳米尺度表征Apobec3G与RNA的相互作用

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摘要

The human cytidine deaminase APOBEC3G (A3G) is a potent inhibitor of the HIV-1 virus in the absence of viral infectivity factor (Vif). The molecular mechanism of A3G antiviral activity is primarily attributed to deamination of single stranded DNA (ssDNA); however, the nondeamination mechanism also contributes to HIV-1 restriction. The interaction of A3G with ssDNA and RNA is required for its antiviral activity. Here we used atomic force microscopy to directly visualize A3G RNA and A3G ssDNA complexes and compare them to each other. Our results showed that A3G in A3G RNA complexes exists primarily in monomeric dimeric states, similar to its stoichiometry in complexes with ssDNA. New A3G RNA complexes in which A3G binds to two RNA molecules were identified. These data suggest the existence of two separate RNA binding sites on A3G. Such complexes were not observed with ssDNA substrates. Time-lapse high-speed atomic force microscopy was applied to characterize the dynamics of the complexes. The data revealed that the two RNA binding sites have different affinities for A3G. On the basis of the obtained results, a model for the interaction of A3G with RNA is proposed.
机译：人胞苷脱氨酶Apobec3G（A3G）是在没有病毒感染因子（VIF）的情况下的HIV-1病毒的有效抑制剂。 A3G抗病毒活性的分子机制主要归因于单链DNA（SSDNA）的核氨酸;然而，非恐机机制也有助于HIV-1限制。 A3G与SSDNA和RNA的相互作用是其抗病毒活性所必需的。在这里，我们使用原子力显微镜直接可视化A3G RNA和A3G SSDNA复合物并将它们彼此进行比较。我们的结果表明，A3G RNA复合物中的A3G主要存在于单体二聚体状态，类似于其与SSDNA复合物的化学计量。新的A3G RNA配合物，其中鉴定了A3G与两个RNA分子结合。这些数据表明A3G上存在两个单独的RNA结合位点。使用SSDNA底物未观察到这种复合物。延时高速原子力显微镜用于表征复合物的动态。数据显示，两个RNA结合位点对A3G具有不同的亲和力。在得到的结果的基础上，提出了一种具有RNA的A3G相互作用的模型。

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  • 来源
    《Biochemistry》 |2017年第10期|共9页
  • 作者

    Pan Yangang; Sun Zhiqiang; Maiti Atanu; Kanai Tapan; Matsuo Hiroshi; Li Ming; Harris Reuben S.; Shlyakhtenko Luda S.; Lyubchenko Yuri L.;

  • 作者单位

    Univ Nebraska Med Ctr WSH Coll Pharm Department Pharmaceut Sci 986025 Nebraska Med Ctr Omaha NE 68198 USA;

    Univ Nebraska Med Ctr WSH Coll Pharm Department Pharmaceut Sci 986025 Nebraska Med Ctr Omaha NE 68198 USA;

    Leidos Biomedical Res Inc Adv Technol Res Facil Frederick Natl Lab Canc Res 8560 Progress Dr Frederick MD 21702 USA;

    Leidos Biomedical Res Inc Adv Technol Res Facil Frederick Natl Lab Canc Res 8560 Progress Dr Frederick MD 21702 USA;

    Leidos Biomedical Res Inc Adv Technol Res Facil Frederick Natl Lab Canc Res 8560 Progress Dr Frederick MD 21702 USA;

    Univ Minnesota Dept Biochem Mol Biol &

    Biophys Inst Mol Virol Ctr Genome Engn Mason Canc Ctr Minneapolis MN 55455 USA;

    Univ Minnesota Dept Biochem Mol Biol &

    Biophys Inst Mol Virol Ctr Genome Engn Mason Canc Ctr Minneapolis MN 55455 USA;

    Univ Nebraska Med Ctr WSH Coll Pharm Department Pharmaceut Sci 986025 Nebraska Med Ctr Omaha NE 68198 USA;

    Univ Nebraska Med Ctr WSH Coll Pharm Department Pharmaceut Sci 986025 Nebraska Med Ctr Omaha NE 68198 USA;

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  • 正文语种 eng
  • 中图分类 生物化学;
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