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Solution structure of the C-terminal domain of the vaccinia virus DNA polymerase processivity factor component A20 fused to a short peptide from the viral DNA polymerase E9.
Authors
Bersch, B., Tarbouriech, N., Burmeister, W., Iseni, F.
Assembly
DNA polymerase processivity factor component A20,DNA polymerase processivity factor component E9
Entity
1. DNA polymerase processivity factor component A20,DNA polymerase processivity factor component E9 (polymer), 151 monomers, 16863.76 Da Detail

GNGKYFSKVG SAGLKQLTNK LDINECATVD ELVDEINKSG TVKRKIKNQS AFDLSRECLG YPEADFITLV NNMRFKIENC KVVNFNIENT NCLNNPSIET IYRNFNQFVS IFNVVTDVKK RLFENASGNG SGGGSNRLEE EINNQLALQK S


Formula weight
16863.76 Da
Source organism
Vaccinia virus Copenhagen
Exptl. method
solution NMR
Data set
assigned_chemical_shifts, heteronucl_NOEs, heteronucl_T1_relaxation, heteronucl_T2_relaxation, spectral_peak_list
Chem. Shift Complete
Sequence coverage: 100.0 %, Completeness: 90.4 %, Completeness (bb): 98.4 % Detail

Polymer type: polypeptide(L)

Total1H13C15N
All90.4 % (1605 of 1776)91.9 % (851 of 926)87.4 % (584 of 668)93.4 % (170 of 182)
Backbone98.4 % (888 of 902)98.1 % (305 of 311)98.4 % (435 of 442)99.3 % (148 of 149)
Sidechain84.3 % (855 of 1014)88.8 % (546 of 615)78.4 % (287 of 366)66.7 % (22 of 33)
Aromatic47.4 % (54 of 114)94.7 % (54 of 57) 0.0 % (0 of 57)
Methyl100.0 % (158 of 158)100.0 % (79 of 79)100.0 % (79 of 79)

1. entity 1

GNGKYFSKVG SAGLKQLTNK LDINECATVD ELVDEINKSG TVKRKIKNQS AFDLSRECLG YPEADFITLV NNMRFKIENC KVVNFNIENT NCLNNPSIET IYRNFNQFVS IFNVVTDVKK RLFENASGNG SGGGSNRLEE EINNQLALQK S

Sample #1

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 (±0.5) K, pH 6 (±0.1), Details 1 mM [U-15N] A20-i3, 50 mM potassium phosphate, 300 mM NaCl, 1.2 mM TCEP, 90% H2O/10% D2O


#NameIsotope labelingTypeConcentration
1A20-i3[U-15N]1 mM
2potassium phosphatenatural abundance50 mM
3NaClnatural abundance300 mM
4TCEPnatural abundance1.2 mM
Sample #2

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 (±0.5) K, pH 6 (±0.1), Details 0.8 mM [U-15N13C] A20-i3, 50 mM potassium phosphate, 300 mM NaCl, 1.2 mM TCEP, 90% H2O/10% D2O


#NameIsotope labelingTypeConcentration
5A20-i3[U-15N; U-13C]0.8 mM
6potassium phosphatenatural abundance50 mM
7NaClnatural abundance300 mM
8TCEPnatural abundance1.2 mM
Sample #3

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 (±0.5) K, pH 6 (±0.1), Details 1 mM [U-15N13C] A20-i3, 50 mM potassium phosphate, 300 mM NaCl, 1.2 mM TCEP, 90% H2O/10% D2O


#NameIsotope labelingTypeConcentration
9A20-i3[U-15N; U-13C]1 mM
10potassium phosphatenatural abundance50 mM
11NaClnatural abundance300 mM
12TCEPnatural abundance1.2 mM

Heteronucl. T1
116 T1 values in 1 lists
Coherence Sz, Field strength (1H) 700.2533 MHz, Pressure 1 atm, Temperature 298 (±0.5) K, pH 6 (±0.1) Detail
Heteronucl. T2
116 T2 values in 1 lists
Coherence S(+,-), Field strength (1H) 700.2533 MHz, Pressure 1 atm, Temperature 298 (±0.5) K, pH 6 (±0.1) Detail
Heteronucl. NOE
118 NOE values in 1 lists
Value type peak height, Field strength (1H) 700 MHz, Pressure 1 atm, Temperature 298 (±0.5) K, pH 6 (±0.1) Detail
Heteronucl. T1/T2
116 T1/T2 values in 1 lists
Field strength (1H) 700.2533 MHz, Pressure 1 atm, Temperature 298 (±0.5) K, pH 6 (±0.1) Detail
Release date
2020-08-27
Citation 1
The vaccinia virus DNA polymerase structure provides insights into the mode of processivity factor binding
Tarbouriech, N., Ducournau, C., Hutin, S., Mas, P.J., Man, P., Forest, E., Hart, D.J., Peyrefitte, C.N., Burmeister, W., Iseni, F.
Nat. Commun. (2017), 8, 1455-1455, PubMed 29129932 , DOI 10.1038/s41467-017-01542-z ,
Citation 2
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
Bersch, B., Tarbouriech, N., Burmeister, W., Iseni, F.
J. Mol. Biol. (2021), 433, 167009-167009, PubMed 33901538 , DOI 10.1016/j.jmb.2021.167009 ,
Related entities 1. DNA polymerase processivity factor component A20,DNA polymerase processivity factor component E9, : 1 : 1 : 3 entities Detail
Interaction partners 1. DNA polymerase processivity factor component A20,DNA polymerase processivity factor component E9, : 4 interactors Detail
Experiments performed 14 experiments Detail
Chemical shift validation 3 contents Detail
Keywords DNA polymerase holoenzyme, Poxviridae, REPLICATION, processivity factor binding