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Full-length human cytomegalovirus terminase pUL89 adopts a two-domain structure specific for DNA packaging.
PLoS Pathogens ( IF 5.5 ) Pub Date : 2019-12-06 , DOI: 10.1371/journal.ppat.1008175 Janine Theiß 1 , Min Woo Sung 2 , Andreas Holzenburg 3 , Elke Bogner 1
PLoS Pathogens ( IF 5.5 ) Pub Date : 2019-12-06 , DOI: 10.1371/journal.ppat.1008175 Janine Theiß 1 , Min Woo Sung 2 , Andreas Holzenburg 3 , Elke Bogner 1
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A key step in replication of human cytomegalovirus (HCMV) in the host cell is the generation and packaging of unit-length genomes into preformed capsids. The enzymes involved in this process are the terminases. The HCMV terminase complex consists of two terminase subunits, the ATPase pUL56 and the nuclease pUL89. A potential third component pUL51 has been proposed. Even though the terminase subunit pUL89 has been shown to be essential for DNA packaging and interaction with pUL56, it is not known how pUL89 mechanistically achieves sequence-specific DNA binding and nicking. To identify essential domains and invariant amino acids vis-a-vis nuclease activity and DNA binding, alanine substitutions of predicted motifs were analyzed. The analyses indicated that aspartate 463 is an invariant amino acid for the nuclease activity, while argine 544 is an invariant aa for DNA binding. Structural analysis of recombinant protein using electron microscopy in conjunction with single particle analysis revealed a curvilinear monomer with two distinct domains connected by a thinner hinge-like region that agrees well with the predicted structure. These results allow us to model how the terminase subunit pUL89's structure may mediate its function.
中文翻译：
全长人巨细胞病毒末端酶pUL89采用DNA包装特有的两个结构域结构。
人巨细胞病毒（HCMV）在宿主细胞中复制的关键步骤是将单位长度基因组生成并包装到预先形成的衣壳中。参与该过程的酶是末端酶。HCMV末端酶复合物由两个末端酶亚基组成，即ATPase pUL56和核酸酶pUL89。已经提出了潜在的第三组分pUL51。尽管已显示末端酶亚基pUL89对于DNA包装和与pUL56相互作用必不可少，但未知pUL89如何以机械方式实现序列特异性DNA结合和切口。为了鉴定相对于核酸酶活性和DNA结合的必需结构域和恒定氨基酸，分析了预测基序的丙氨酸取代。分析表明，天冬氨酸463是核酸酶活性的不变氨基酸，而精氨酸544是DNA结合的恒定氨基酸。使用电子显微镜结合单颗粒分析对重组蛋白进行结构分析，结果显示出一个曲线型单体，具有两个不同的结构域，这些结构域由较薄的类似于预测结构的铰链状区域连接。这些结果使我们能够对末端酶亚单位pUL89的结构如何介导其功能进行建模。
更新日期：2019-12-07
中文翻译：
全长人巨细胞病毒末端酶pUL89采用DNA包装特有的两个结构域结构。
人巨细胞病毒（HCMV）在宿主细胞中复制的关键步骤是将单位长度基因组生成并包装到预先形成的衣壳中。参与该过程的酶是末端酶。HCMV末端酶复合物由两个末端酶亚基组成，即ATPase pUL56和核酸酶pUL89。已经提出了潜在的第三组分pUL51。尽管已显示末端酶亚基pUL89对于DNA包装和与pUL56相互作用必不可少，但未知pUL89如何以机械方式实现序列特异性DNA结合和切口。为了鉴定相对于核酸酶活性和DNA结合的必需结构域和恒定氨基酸，分析了预测基序的丙氨酸取代。分析表明，天冬氨酸463是核酸酶活性的不变氨基酸，而精氨酸544是DNA结合的恒定氨基酸。使用电子显微镜结合单颗粒分析对重组蛋白进行结构分析，结果显示出一个曲线型单体，具有两个不同的结构域，这些结构域由较薄的类似于预测结构的铰链状区域连接。这些结果使我们能够对末端酶亚单位pUL89的结构如何介导其功能进行建模。
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