Npl3 functions in mRNP assembly by recruitment of mRNP components to the transcription site and their transfer onto the mRNA.

Keil P, Wulf A, Kachariya N Reuscher S, Hühn K, Silbern I, Altmüller J, Keller M, Stehle R, Zarnack K, Sattler M, Urlaub H, Sträßer K, Nucleic Acids Res (2022) Europe PMC

SASDQV5 – NPL3 protein RNA recognition motifs 1 and 2 (RRM1,2) wild-type, bound to RNA

Serine/arginine (SR)-type shuttling mRNA binding protein NPL3
Short RNA oligonucleotide (NPL3 binding sequence)
MWexperimental 23 kDa
MWexpected 22 kDa
VPorod 30 nm3
log I(s) 9.02×10-1 9.02×10-2 9.02×10-3 9.02×10-4
Serine/arginine (SR)-type shuttling mRNA binding protein NPL3 Short RNA oligonucleotide (NPL3 binding sequence) small angle scattering data  s, nm-1
ln I(s)
Serine/arginine (SR)-type shuttling mRNA binding protein NPL3 Short RNA oligonucleotide (NPL3 binding sequence) Guinier plot ln 9.02×10-1 Rg: 2 nm 0 (2 nm)-2 s2
(sRg)2I(s)/I(0)
Serine/arginine (SR)-type shuttling mRNA binding protein NPL3 Short RNA oligonucleotide (NPL3 binding sequence) Kratky plot 1.104 0 √3 sRg
p(r)
Serine/arginine (SR)-type shuttling mRNA binding protein NPL3 Short RNA oligonucleotide (NPL3 binding sequence) pair distance distribution function Rg: 2.1 nm 0 Dmax: 6.3 nm

Data validation

There are no models related to this curve.

SAXS data from solutions of the NPL3 protein RNA recognition motif domains 1 and 2 (RRM1,2) bound to RNA in 20 mM NaPO4, 50 mM NaCl, 1 mM DTT, pH 6.5 were collected using a Rigaku bioSAXS-1000 instrument at the Technische Universität München (TUM; Garching, Germany) equipped with a Pilatus 100K detector at a sample-detector distance of 0.5 m and at a wavelength of λ = 0.155 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 4.90 mg/ml was measured at 25°C. Eight successive 900 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 RNA bound NPL3 complex was prepared by mixing the equimolar ratio of protein to RNA. The complex was further purified using gel-filtration chromatography. The short RNA sequence 5'-AGCACCGUGGAGA-3' was used in preparation of protein-RNA complex.

Serine/arginine (SR)-type shuttling mRNA binding protein NPL3
Mol. type   Protein
Organism   Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Olig. state   Monomer
Mon. MW   18.2 kDa
 
UniProt   Q01560 (120-280)
Sequence   FASTA
 
Short RNA oligonucleotide (NPL3 binding sequence)
Mol. type   RNA
Olig. state   Monomer
Mon. MW   4.3 kDa
Sequence   FASTA