Subdomain specific functions of the RNA polymerase region of poliovirus 3CD polypeptide
Subdomain specific functions of the RNA polymerase region of poliovirus 3CD polypeptide
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DOI：
10.1006/viro.2002.1481
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发表时间：
2002-07-05
期刊：
影响因子：
3.7
通讯作者：
Semler, BL
中科院分区：
文献类型：
--
作者：
Cornell, CT;Semler, BL
脊髓灰质炎病毒3CD多肽的3D聚合酶结构域在调节其RNA结合和蛋白质加工活性方面发挥作用，尽管蛋白酶催化位点和RNA结合决定簇似乎位于分子的3C(Pro)部分。在这项研究中，我们产生了含有嵌合3D聚合酶结构域的重组KID多肽，这些嵌合3D聚合酶结构域代表了脊髓灰质炎病毒1型(PV1)和柯萨奇病毒B3(CVB3)之间的亚等位基因序列交换，以确定3D结构域的哪些部分负责影响这些活性。通过将这些重组蛋白嵌合体用于体外蛋白质加工和RNA结合研究，我们产生的数据表明，3CD的聚合酶结构域中存在单独的亚域，这些亚域可能独立地调节其RNA结合和蛋白质加工活性。在利用先前发表的PV1 3D聚合酶的三维结构预测我们的序列交换映射的位置时，我们提供了证据，表明该聚合酶的RNA识别基序中包含的序列对3CD功能在识别5‘RNA三叶草中起关键作用。此外，我们的蛋白质加工数据表明，3CD的3D结构域中至少有一些底物识别和加工决定因素与该区域的RNA结合决定因素是分开和不同的。(C)2002年埃尔塞维尔科学公司(美国)。
The 3D polymerase domain of the poliovirus 3CD polypeptide plays a role in modulating its RNA binding and protein processing activities, even though the proteinase catalytic site and RNA binding determinants appear to reside within the 3C(pro) portion of the molecule. In this study, we have generated recombinant KID polypeptides that contain chimeric 3D polymerase domains representing suballelic sequence exchanges between poliovirus type 1 (PV1) and coxsackievirus B3 (CVB3) to determine which portions of the 3D domain are responsible for influencing these activities. By utilizing these recombinant protein chimeras in protein processing and RNA binding studies in vitro, we have generated data suggesting the presence of separate subdomains within the polymerase domain of 3CD that may independently modulate its RNA binding and protein processing activities. In predicting where our sequence exchanges map by utilizing the previously published three-dimensional structure of the PV1 3D polymerase, we present evidence that sequences contained within the RNA recognition motif of the polymerase are critical for 3CD function in recognizing the 5' RNA cloverleaf. Furthermore, our protein processing data indicate that at least some of the substrate recognition and processing determinants within the 3D domain of 3CD are separate and distinct from the RNA binding determinants in this domain. (C) 2002 Elsevier Science (USA).