Solution Structure of the C-terminal Domain of A20, the Missing Brick for the Characterization of the Interface between Vaccinia Virus DNA Polymerase and its Processivity Factor
Copyright © 2021 Elsevier Ltd. All rights reserved..
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 A20304-426-E9 complex, which is in excellent accordance with previous experimental data.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:433 |
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| Enthalten in: |
Journal of molecular biology - 433(2021), 13 vom: 25. Juni, Seite 167009 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Bersch, Beate [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 30.08.2021 Date Revised 30.08.2021 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.jmb.2021.167009 |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM324550316 |
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| 245 | 1 | 0 | |a Solution Structure of the C-terminal Domain of A20, the Missing Brick for the Characterization of the Interface between Vaccinia Virus DNA Polymerase and its Processivity Factor |
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| 500 | |a Date Completed 30.08.2021 | ||
| 500 | |a Date Revised 30.08.2021 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2021 Elsevier Ltd. All rights reserved. | ||
| 520 | |a 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 A20304-426-E9 complex, which is in excellent accordance with previous experimental data | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a DNA replication | |
| 650 | 4 | |a NMR | |
| 650 | 4 | |a holoenzyme | |
| 650 | 4 | |a poxvirus | |
| 650 | 4 | |a protein-protein interaction | |
| 650 | 7 | |a DNA, Viral |2 NLM | |
| 650 | 7 | |a Holoenzymes |2 NLM | |
| 650 | 7 | |a Peptides |2 NLM | |
| 650 | 7 | |a Solutions |2 NLM | |
| 650 | 7 | |a Viral Proteins |2 NLM | |
| 650 | 7 | |a DNA polymerase, vaccinia virus |2 NLM | |
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| 650 | 7 | |a DNA-Directed DNA Polymerase |2 NLM | |
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| 700 | 1 | |a Tarbouriech, Nicolas |e verfasserin |4 aut | |
| 700 | 1 | |a Burmeister, Wim P |e verfasserin |4 aut | |
| 700 | 1 | |a Iseni, Frédéric |e verfasserin |4 aut | |
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