BMRB Entry 28090
Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR28090
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Title: 1H, 13C, and 15N chemical shifts for T4 Gene 60 mRNA 5' Stem-Loop PubMed: 32454154
Deposition date: 2020-02-29 Original release date: 2021-07-16
Authors: Capece, Mark; Puglisi, Joseph
Citation: O'Loughlin, Sinead; Capece, Mark; Klimova, Mariia; Wills, Norma; Coakley, Arthur; Samatova, Ekaterina; O'Connor, Patrick; Loughran, Gary; Weissman, Jonathan; Baranov, Pavel; Rodnina, Marina; Puglisi, Joseph; Atkins, John. "Polysomes bypass a 50 nucleotide coding gap less efficiently than monosomes due to attenuation of an unstable 5' mRNA stem loop stimulator and enhanced drop-off" J. Mol. Biol. 432, 4369-4387 (2020).
Assembly members:
5'_stem_loop, polymer, 30 residues, Formula weight is not available
Natural source: Common Name: T4 bacteriophage Taxonomy ID: 38018 Superkingdom: Viruses Kingdom: not available Genus/species: T4 bacteriophage
Experimental source: Production method: in vitro transcription Host organism: T7 RNA polymerase Vector: N/A
Entity Sequences (FASTA):
5'_stem_loop: GUGCGAAUGUCGCUAUUAUG
ACAGACGCAC
- assigned_chemical_shifts
| Data type | Count |
| 13C chemical shifts | 73 |
| 15N chemical shifts | 44 |
| 1H chemical shifts | 135 |
Additional metadata:
Assembly:
| Entity Assembly ID | Entity Name | Entity ID |
|---|---|---|
| 1 | 5' stem loop | 1 |
Entities:
Entity 1, 5' stem loop 30 residues - Formula weight is not available
| 1 | G | U | G | C | G | A | A | U | G | U | |
| 2 | C | G | C | U | A | U | U | A | U | G | |
| 3 | A | C | A | G | A | C | G | C | A | C |
Samples:
10%_D2O: 5' stem loop, [U-99% 13C; U-99% 15N], 0.3 mM; sodium phosphate 10 mM; sodium chloride 20 mM
99.99%_D2O: 5' stem loop, [U-99% 13C; U-99% 15N], 0.3 mM; sodium phosphate 10 mM; sodium chloride 20 mM
10%_D2O_conditions: pH: 6.5; pressure: 1 atm; temperature: 298 K
99.99%_D2O_conditions: pH: 6.5; pressure: 1 atm; temperature: 298 K
Experiments:
| Name | Sample | Sample state | Sample conditions |
|---|---|---|---|
| 2D 1H-1H NOESY | 10%_D2O | isotropic | 10%_D2O_conditions |
| 2D 1H-15N HSQC | 10%_D2O | isotropic | 10%_D2O_conditions |
| 2D 1H-15N HNNCOSY | 10%_D2O | isotropic | 10%_D2O_conditions |
| 3D 1H-15N NOESYHSQC | 10%_D2O | isotropic | 10%_D2O_conditions |
| 3D HNCCCH | 10%_D2O | isotropic | 10%_D2O_conditions |
| 2D 1H-1H NOESY | 99.99%_D2O | isotropic | 99.99%_D2O_conditions |
| 2D 1H-13C HSQC | 99.99%_D2O | isotropic | 99.99%_D2O_conditions |
| 3D 1H-13C NOESYHSQC | 99.99%_D2O | isotropic | 99.99%_D2O_conditions |
| 3D HCCH-TOCSY | 99.99%_D2O | isotropic | 99.99%_D2O_conditions |
| 2D DQF-COSY | 99.99%_D2O | isotropic | 99.99%_D2O_conditions |
Software:
VNMRJ v4.2, Varian - collection, processing
SPARKY, Goddard - chemical shift assignment, peak picking
NMR spectrometers:
- Agilent VNMRS 800 MHz
- Agilent INOVA 600 MHz