Despite the odds: formation of the SARS-CoV-2 methylation complex

Matsuda A, Plewka J, Rawski M, Mourão A, Zajko W, Siebenmorgen T, Kresik L, Lis K, Jones A, Pachota M, Karim A, Hartman K, Nirwal S, Sonani R, Chykunova Y, Minia I, Mak P, Landthaler M, Nowotny M, Dubin G, Sattler M, Suder P, Popowicz G, Pyrć K, Czarna A, Nucleic Acids Research (2024) DOI

SASDKT6 – SARS-CoV-2 non-structural protein 14 (nsp14)

Replicase polyprotein 1ab (non-structural protein 14)
MWexperimental 33 kDa
MWexpected 60 kDa
VPorod 80 nm3
log I(s) 8.22×100 8.22×10-1 8.22×10-2 8.22×10-3
Replicase polyprotein 1ab (non-structural protein 14) small angle scattering data  s, nm-1
ln I(s)
Replicase polyprotein 1ab (non-structural protein 14) Guinier plot ln 8.22×100 Rg: 2.7 nm 0 (2.7 nm)-2 s2
(sRg)2I(s)/I(0)
Replicase polyprotein 1ab (non-structural protein 14) Kratky plot 1.104 0 3 sRg
p(r)
Replicase polyprotein 1ab (non-structural protein 14) pair distance distribution function Rg: 2.9 nm 0 Dmax: 9.6 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Replicase polyprotein 1ab (non-structural protein 14) PYMOL model

log I(s)
 s, nm-1
Replicase polyprotein 1ab (non-structural protein 14) GASBOR model

Synchrotron SAXS data from solutions of SARS-CoV-2 non-structural protein 14 (nsp14) in 50 mM Tris, 150 mM NaCl, 5 mM MgCl2, 2 mM β-mercaptoethanol, pH 8.5 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.8 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 100.00 μl sample at 4 mg/ml was injected onto a Agilent Bio SEC-3, 300 Å column. 21 successive frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

CAUTION! A significant discrepancy is noted between the experimentally determined (33 kDa) vs. expected (60 kDa) molecular weights. Experimental temperature: UNKNOWN. X-ray exposure time: UNKNOWN. SEC-flow rate: UNKNOWN.

Replicase polyprotein 1ab (non-structural protein 14) (nsp14)
Mol. type   Protein
Organism   Severe acute respiratory syndrome coronavirus 2
Olig. state   Monomer
Mon. MW   60.1 kDa
 
UniProt   P0DTD1 (5926-6452)
Sequence   FASTA
 
PDB ID   5C8U