Short prokaryotic Argonautes provide defence against incoming mobile genetic elements through NAD+ depletion

Zaremba M, Dakineviciene D, Golovinas E, Zagorskaitė E, Stankunas E, Lopatina A, Sorek R, Manakova E, Ruksenaite A, Silanskas A, Asmontas S, Grybauskas A, Tylenyte U, Jurgelaitis E, Grigaitis R, Timinskas K, Venclovas Č, Siksnys V, Nature Microbiology (2022) DOI

SASDNH2 – Complex of Geobacter sulfurreducens pAgo with Sir2-like effector protein

Piwi domain protein
Sir2 superfamily protein
MWI(0) 101 kDa
MWexpected 121 kDa
VPorod 185 nm3
log I(s) 7.34×10-2 7.34×10-3 7.34×10-4 7.34×10-5
Piwi domain protein Sir2 superfamily protein small angle scattering data  s, nm-1
ln I(s)
Piwi domain protein Sir2 superfamily protein Guinier plot ln 7.34×10-2 Rg: 3.6 nm 0 (3.6 nm)-2 s2
(sRg)2I(s)/I(0)
Piwi domain protein Sir2 superfamily protein Kratky plot 1.104 0 √3 sRg
p(r)
Piwi domain protein Sir2 superfamily protein pair distance distribution function Rg: 3.6 nm 0 Dmax: 11.4 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Piwi domain protein Sir2 superfamily protein GASBOR model
Synchrotron SAXS data from solutions of the complex formed between Geobacter sulfurreducens pAgo with Sir2-like effector protein in 20 mM TrisHCl, pH 7.5, 5 mM MgCl2, 150 mM NaCl and 2 mM DTT, pH 7.5 were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 1.2 and 6.3 mg/ml were measured at 20°C. 20 successive 0.045 second 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. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Data describe the stable complex between G. sulfurreducens Argonaute and Sir2-like effector protein. The complex was purified by gel filtration prior to measurement.

Piwi domain protein
Mol. type   Protein
Organism   Geobacter sulfurreducens (strain ATCC 51573 / DSM 12127 / PCA)
Olig. state   Monomer
Mon. MW   53.3 kDa
 
UniProt   Q74DF5 (1-473)
Sequence   FASTA
 
Sir2 superfamily protein (Sir2)
Mol. type   Protein
Organism   Geobacter sulfurreducens (strain ATCC 51573 / DSM 12127 / PCA)
Olig. state   Monomer
Mon. MW   68 kDa
 
UniProt   Q74DF6 (1-587)
Sequence   FASTA