A possible role for the asymmetric C-terminal domain dimer of Rous sarcoma virus integrase in viral DNA binding.
A possible role for the asymmetric C-terminal domain dimer of Rous sarcoma virus integrase in viral DNA binding.
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DOI：
10.1371/journal.pone.0056892
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Aihara H
中科院分区：
文献类型：
--
作者：
Shi K;Pandey KK;Bera S;Vora AC;Grandgenett DP;Aihara H
逆转录病毒线性 DNA 基因组整合到宿主染色体中是病毒复制周期中的重要步骤，并由病毒整合酶 (IN) 催化。有证据表明，IN 作为二聚体发挥作用，从 3' DNA 平端切割二核苷酸，而二聚体的二聚体（四聚体）则促进两个加工末端协同整合到目标 DNA 的相反链中。然而，目前尚不清楚为什么插入每个凹陷的 DNA 末端需要 IN 二聚体而不是单体。为了帮助解决这个问题，我们以更高的分辨率分析了劳斯肉瘤病毒 (RSV) IN 突变体的晶体结构，其中包括所有三个结构域以及新晶体形式的两个结构域片段。结合早期的结构研究，我们的结果表明，RSV IN 二聚体由高度灵活的 N 端结构域和由催化结构域和 C 端结构域形成的刚性实体组成，该结构域通过良好保守的催化结构域二聚相互作用稳定。生化和突变分析证实了早期的观察结果，即 RSV IN 二聚体的催化和 C 末端结构域有效地将一个病毒 DNA 末端整合到目标 DNA 中。我们还表明，两个 C 端结构域之间的不对称二聚体相互作用对于病毒 DNA 结合和随后的催化（包括协同整合）非常重要。我们建议不对称 C 端结构域二聚体作为 RSV IN 的病毒 DNA 结合表面。
Integration of the retrovirus linear DNA genome into the host chromosome is an essential step in the viral replication cycle, and is catalyzed by the viral integrase (IN). Evidence suggests that IN functions as a dimer that cleaves a dinucleotide from the 3′ DNA blunt ends while a dimer of dimers (tetramer) promotes concerted integration of the two processed ends into opposite strands of a target DNA. However, it remains unclear why a dimer rather than a monomer of IN is required for the insertion of each recessed DNA end. To help address this question, we have analyzed crystal structures of the Rous sarcoma virus (RSV) IN mutants complete with all three structural domains as well as its two-domain fragment in a new crystal form at an improved resolution. Combined with earlier structural studies, our results suggest that the RSV IN dimer consists of highly flexible N-terminal domains and a rigid entity formed by the catalytic and C-terminal domains stabilized by the well-conserved catalytic domain dimerization interaction. Biochemical and mutational analyses confirm earlier observations that the catalytic and the C-terminal domains of an RSV IN dimer efficiently integrates one viral DNA end into target DNA. We also show that the asymmetric dimeric interaction between the two C-terminal domains is important for viral DNA binding and subsequent catalysis, including concerted integration. We propose that the asymmetric C-terminal domain dimer serves as a viral DNA binding surface for RSV IN.
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影响因子：
4.8
作者：
Jenkins, TM;Engelman, A;Craigie, R
通讯作者：
Craigie, R
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
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作者：
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通讯作者：
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DOI：
10.1073/pnas.150220297
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影响因子：
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通讯作者：
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DOI：
10.1038/nsb0995-807
发表时间：
1995-09-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
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作者：
EIJKELENBOOM, APAM;LUTZKE, RAP;HARD, K
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影响因子：
14.9
作者：
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通讯作者：
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