The molecular dissection of TRIM25's RNA-binding mechanism provides key insights into its antiviral activity.

Álvarez L, Haubrich K, Iselin L, Gillioz L, Ruscica V, Lapouge K, Augsten S, Huppertz I, Choudhury NR, Simon B, Masiewicz P, Lethier M, Cusack S, Rittinger K, Gabel F, Leitner A, Michlewski G, Hentze MW, Allain FHT, Castello A, Hennig J, Nat Commun 15(1):8485 (2024) Europe PMC

SASDK98 – E3 ubiquitin/ISG15 ligase TRIM25 bound to pre-let-7-a-1@1 RNA (TRIM25/pre-let-7): SEC-SAXS

E3 ubiquitin/ISG15 ligase TRIM25
pre-let-7-a-1@1
MWexperimental 109 kDa
MWexpected 109 kDa
log I(s) 2.16×101 2.16×100 2.16×10-1 2.16×10-2
E3 ubiquitin/ISG15 ligase TRIM25 pre-let-7-a-1@1 small angle scattering data  s, nm-1
ln I(s)
E3 ubiquitin/ISG15 ligase TRIM25 pre-let-7-a-1@1 Guinier plot ln 2.17×101 Rg: 5.7 nm 0 (5.7 nm)-2 s2
(sRg)2I(s)/I(0)
E3 ubiquitin/ISG15 ligase TRIM25 pre-let-7-a-1@1 Kratky plot 1.104 0 √3 sRg
Dmax: 23 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of TRIM25/pre-let-7 in 20 mM MES, 75 mM NaCl, 1 mM TCEP, pH 6.5 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.099 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 sample was injected onto a GE Superdex 200 Increase 10/300 column at 20°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Sample injection concentration: UNKNOWN. Sample injection volume: UNKNOWN. SEC column flow rate: UNKNOWN. Total X-ray exposure time and number of individual SEC-SAXS data frames used for processing: UNKNOWN.

E3 ubiquitin/ISG15 ligase TRIM25 (TRIM25)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   50.1 kDa
 
UniProt   Q14258 (189-630)
Sequence   FASTA
 
pre-let-7-a-1@1 (pre-let-7)
Mol. type   RNA
Organism   synthetic construct
Olig. state   Monomer
Mon. MW   8.9 kDa
Sequence   FASTA