期刊
JOURNAL OF MOLECULAR BIOLOGY
卷 433, 期 13, 页码 -出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2021.167009
关键词
NMR; protein-protein interaction; poxvirus; DNA replication; holoenzyme
资金
- Service de Sante des Armees
- French National Research Agency [ANR-15-IDEX-02]
- Fond de la Recherche Medicale [DEQ20170336754]
- TGIR-RMN-THC Fr3050 CNRS
- FRISBI [ANR-10-INBS-0005-02]
- GRAL
- University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS [ANR-17-EURE-0003]
- Direction Generale pour l'Armement
Poxviruses are enveloped viruses with a linear, double-stranded DNA genome, and viral DNA synthesis is achieved by a DNA polymerase holoenzyme composed of three essential proteins. The catalytic subunit E9 forms a complex with the heterodimeric processivity factor made up of D4 and A20, which have a specific interaction involving a poxvirus-specific insertion. The interface between E9 and A20 comprises conserved hydrophobic residues and has been structurally characterized, providing insights into their molecular interactions.
Poxviruses are enveloped viruses with a linear, double-stranded DNA genome. Viral DNA synthesis is achieved by a functional DNA polymerase holoenzyme composed of three essential proteins. For vaccinia virus (VACV) these are E9, the catalytic subunit, a family B DNA polymerase, and the heterodimeric processivity factor formed by D4 and A20. The A20 protein links D4 to the catalytic subunit. High-resolution structures have been obtained for the VACV D4 protein in complex with an N-terminal fragment of A20 as well as for E9. In addition, biochemical studies provided evidence that a poxvirus-specific insertion (insert 3) in E9 interacts with the C-terminal residues of A20. Here, we provide solution structures of two different VACV A20 C-terminal constructs containing residues 304-426, fused at their C-terminus to either a BAP (Biotin Acceptor Peptide)-tag or a short peptide containing the helix of E9 insert 3. Together with results from titration studies, these structures shed light on the molecular interface between the catalytic subunit and the processivity factor component A20. The interface comprises hydrophobic residues conserved within the Chordopoxvirinae subfamily. Finally, we constructed a HADDOCK model of the VACV A20(304-426)-E9 complex, which is in excellent accordance with previous experimental data. (C) 2021 Elsevier Ltd. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据