7NBV

Structure of 2A protein from Theilers murine encephalomyelitis virus (TMEV)

Summary for 7NBV

• Entry DOI: 10.2210/pdb7nbv/pdb
• Descriptor: Capsid protein VP0, BROMIDE ION (3 entities in total)
• Functional Keywords: tmev, prf, recoding, frameshifting, protein-mediated frameshifting, rna-binding protein, viral protein, translational regulation, ribosome-binding
• Biological source: Theiler's murine encephalomyeltits virus GDVII
• Total number of polymer chains: 1
• Total formula weight: 16045.20

Authors

Hill, C.H.,Cook, G.M.,Napthine, S.,Kibe, A.,Brown, K.,Caliskan, N.,Firth, A.E.,Graham, S.C.,Brierley, I. (deposition date: 2021-01-28, release date: 2021-12-08, Last modification date: 2024-06-19)

Primary citation

Hill, C.H.,Cook, G.M.,Napthine, S.,Kibe, A.,Brown, K.,Caliskan, N.,Firth, A.E.,Graham, S.C.,Brierley, I.
Investigating molecular mechanisms of 2A-stimulated ribosomal pausing and frameshifting in Theilovirus.
Nucleic Acids Res., 49:11938-11958, 2021

PubMed Abstract: The 2A protein of Theiler's murine encephalomyelitis virus (TMEV) acts as a switch to stimulate programmed -1 ribosomal frameshifting (PRF) during infection. Here, we present the X-ray crystal structure of TMEV 2A and define how it recognises the stimulatory RNA element. We demonstrate a critical role for bases upstream of the originally predicted stem-loop, providing evidence for a pseudoknot-like conformation and suggesting that the recognition of this pseudoknot by beta-shell proteins is a conserved feature in cardioviruses. Through examination of PRF in TMEV-infected cells by ribosome profiling, we identify a series of ribosomal pauses around the site of PRF induced by the 2A-pseudoknot complex. Careful normalisation of ribosomal profiling data with a 2A knockout virus facilitated the identification, through disome analysis, of ribosome stacking at the TMEV frameshifting signal. These experiments provide unparalleled detail of the molecular mechanisms underpinning Theilovirus protein-stimulated frameshifting.

PubMed: 34751406
DOI: 10.1093/nar/gkab969

Experimental method

X-RAY DIFFRACTION (1.87 Å)