Mechanistic insights into RNA surveillance by the canonical poly(A) polymerase Pla1 of the MTREC complex.

Soni K, Sivadas A, Horvath A, Dobrev N, Hayashi R, Kiss L, Simon B, Wild K, Sinning I, Fischer T, Nat Commun 14(1):772 (2023) Europe PMC

SASDKE6 – Poly(A) polymerase pla1 bound to the NURS complex subunit red1

NURS complex subunit red1
Poly(A) polymerase pla1
MWexperimental 86 kDa
MWexpected 73 kDa
VPorod 138 nm3
log I(s) 1.92×104 1.92×103 1.92×102 1.92×101
NURS complex subunit red1 Poly(A) polymerase pla1 small angle scattering data  s, nm-1
ln I(s)
NURS complex subunit red1 Poly(A) polymerase pla1 Guinier plot ln 1.93×104 Rg: 3.3 nm 0 (3.3 nm)-2 s2
(sRg)2I(s)/I(0)
NURS complex subunit red1 Poly(A) polymerase pla1 Kratky plot 1.104 0 √3 sRg
p(r)
NURS complex subunit red1 Poly(A) polymerase pla1 pair distance distribution function Rg: 3.3 nm 0 Dmax: 10.7 nm

Data validation

Fits and models

log I(s)
 s, nm-1
NURS complex subunit red1 Poly(A) polymerase pla1 CORAL model
Synchrotron SAXS data from solutions the pla1-red1 complex in 20 mM HEPES, pH 7.5, 150 mM NaCl, 1mM DTT, 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 = 0.124193 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 0.85 mg/ml was measured at 20°C. 40 successive 0.195 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.

NURS complex subunit red1 (Red1)
Mol. type   Protein
Organism   Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Olig. state   Monomer
Mon. MW   7.6 kDa
 
UniProt   Q9UTR8 (288-345)
Sequence   FASTA
 
Poly(A) polymerase pla1 (Pla1)
Mol. type   Protein
Organism   Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Olig. state   Monomer
Mon. MW   65.3 kDa
 
UniProt   Q10295 (1-566)
Sequence   FASTA