1887

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Volume 82, Issue 5

The ‘cleavage’ activities of foot-and-mouth disease virus 2A site-directed mutants and naturally occurring ‘2A-like’ sequences Free

Abstract

The 2A/2B cleavage of aphtho- and cardiovirus 2A polyproteins is mediated by their 2A proteins ‘cleaving’ at their own C termini. We have analysed this activity using artificial reporter polyprotein systems comprising green fluorescent protein (GFP) linked via foot-and-mouth disease virus (FMDV) 2A to β-glucuronidase (GUS) – forming a single, long, open reading frame. Analysis of the distribution of radiolabel showed a high proportion of the in translation products (∼90%) were in the form of the ‘cleavage’ products GUS and [GFP2A]. Alternative models have been proposed to account for the ‘cleavage’ activity: proteolysis by a host-cell proteinase, autoproteolysis or a translational effect. To investigate the mechanism of this cleavage event constructs encoding site-directed mutant and naturally occurring ‘2A-like’ sequences were used to program in translation systems and the gel profiles analysed. Analysis of site-directed mutant 2A sequences showed that ‘cleavage’ occurred in constructs in which all the candidate nucleophilic residues were substituted – with the exception of aspartate-12. This residue is not, however, conserved amongst all functional ‘2A-like’ sequences. ‘2A-like’ sequences were identified within insect virus polyproteins, the NS34 protein of type C rotaviruses, repeated sequences in spp. and a eubacterial α-glucosiduronasesequence(). All of the 2A-like sequences analysed were active (to various extents), other than the eubacterial α-glucosiduronase 2A-like sequence. This method of control of protein biogenesis may well not, therefore, be confined to members of the . Taken together, these data provide additional evidence that neither FMDV 2A nor ‘2A-like’ sequences are autoproteolytic elements.

  • Received:
  • Accepted:
  • Published Online:
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2001-05-01
2024-04-19
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The ‘cleavage’ activities of foot-and-mouth disease virus 2A site-directed mutants and naturally occurring ‘2A-like’ sequences
J. Gen. Virol. 82, 1027 (2001); https://doi.org/10.1099/0022-1317-82-5-1027
/content/journal/jgv/10.1099/0022-1317-82-5-1027
/content/journal/jgv/10.1099/0022-1317-82-5-1027
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