Foot-and-mouth disease virus 2A oligopeptide mediated cleavage of an artificial polyprotein - PubMed (original) (raw)

Foot-and-mouth disease virus 2A oligopeptide mediated cleavage of an artificial polyprotein

M D Ryan et al. EMBO J. 1994.

Abstract

We describe the construction of a plasmid (pCAT2AGUS) encoding a polyprotein in which a 19 amino acid sequence spanning the 2A region of the foot-and-mouth disease virus (FMDV) polyprotein was inserted between the reporter genes chloramphenicol acetyl transferase (CAT) and beta-glucuronidase (GUS) maintaining a single, long open reading frame. Analysis of translation reactions programmed by this construct showed that the inserted FMDV sequence functioned in a manner similar to that observed in FMDV polyprotein processing: the CAT2AGUS polyprotein underwent a cotranslational, apparently autoproteolytic, cleavage yielding CAT-2A and GUS. Analysis of translation products derived from a series of constructs in which sequences were progressively deleted from the N-terminal region of the FMDV 2A insertion showed that cleavage required a minimum of 13 residues. The FMDV 2A sequence therefore provides the opportunity to engineer either whole proteins or domains such that they are cleaved apart cotranslationally with high efficiency.

PubMed Disclaimer

Similar articles

• The cleavage activities of aphthovirus and cardiovirus 2A proteins.
  Donnelly ML, Gani D, Flint M, Monaghan S, Ryan MD. Donnelly ML, et al. J Gen Virol. 1997 Jan;78 ( Pt 1):13-21. doi: 10.1099/0022-1317-78-1-13. J Gen Virol. 1997. PMID: 9010280
• Analysis of the aphthovirus 2A/2B polyprotein 'cleavage' mechanism indicates not a proteolytic reaction, but a novel translational effect: a putative ribosomal 'skip'.
  Donnelly MLL, Luke G, Mehrotra A, Li X, Hughes LE, Gani D, Ryan MD. Donnelly MLL, et al. J Gen Virol. 2001 May;82(Pt 5):1013-1025. doi: 10.1099/0022-1317-82-5-1013. J Gen Virol. 2001. PMID: 11297676
• Cleavage of foot-and-mouth disease virus polyprotein is mediated by residues located within a 19 amino acid sequence.
  Ryan MD, King AM, Thomas GP. Ryan MD, et al. J Gen Virol. 1991 Nov;72 ( Pt 11):2727-32. doi: 10.1099/0022-1317-72-11-2727. J Gen Virol. 1991. PMID: 1658199
• Modifications to the Foot-and-Mouth Disease Virus 2A Peptide: Influence on Polyprotein Processing and Virus Replication.
  Kjær J, Belsham GJ. Kjær J, et al. J Virol. 2018 Mar 28;92(8):e02218-17. doi: 10.1128/JVI.02218-17. Print 2018 Apr 15. J Virol. 2018. PMID: 29386286 Free PMC article.
• The 'cleavage' activities of foot-and-mouth disease virus 2A site-directed mutants and naturally occurring '2A-like' sequences.
  Donnelly MLL, Hughes LE, Luke G, Mendoza H, Ten Dam E, Gani D, Ryan MD. Donnelly MLL, et al. J Gen Virol. 2001 May;82(Pt 5):1027-1041. doi: 10.1099/0022-1317-82-5-1027. J Gen Virol. 2001. PMID: 11297677

Cited by

• APE-type non-LTR retrotransposons of multicellular organisms encode virus-like 2A oligopeptide sequences, which mediate translational recoding during protein synthesis.
  Odon V, Luke GA, Roulston C, de Felipe P, Ruan L, Escuin-Ordinas H, Brown JD, Ryan MD, Sukhodub A. Odon V, et al. Mol Biol Evol. 2013 Aug;30(8):1955-65. doi: 10.1093/molbev/mst102. Epub 2013 May 31. Mol Biol Evol. 2013. PMID: 23728794 Free PMC article.
• '2A-Like' Signal Sequences Mediating Translational Recoding: A Novel Form of Dual Protein Targeting.
  Roulston C, Luke GA, de Felipe P, Ruan L, Cope J, Nicholson J, Sukhodub A, Tilsner J, Ryan MD. Roulston C, et al. Traffic. 2016 Aug;17(8):923-39. doi: 10.1111/tra.12411. Epub 2016 Jun 2. Traffic. 2016. PMID: 27161495 Free PMC article.
• Towards optimising the production of and expression from polycistronic vectors in embryonic stem cells.
  Gao SY, Jack MM, O'Neill C. Gao SY, et al. PLoS One. 2012;7(11):e48668. doi: 10.1371/journal.pone.0048668. Epub 2012 Nov 6. PLoS One. 2012. PMID: 23139810 Free PMC article.
• Targeted knock-in of NCF1 cDNA into the NCF2 locus leads to myeloid phenotypic correction of p47 phox -deficient chronic granulomatous disease.
  Siow KM, Güngör M, Wrona D, Raimondi F, Pastukhov O, Tsapogas P, Menzi T, Schmitz M, Kulcsár PI, Schwank G, Schulz A, Jinek M, Modlich U, Siler U, Reichenbach J. Siow KM, et al. Mol Ther Nucleic Acids. 2024 May 24;35(3):102229. doi: 10.1016/j.omtn.2024.102229. eCollection 2024 Sep 10. Mol Ther Nucleic Acids. 2024. PMID: 38952440 Free PMC article.
• Enabling technologies for manipulating multiple genes on complex pathways.
  Halpin C, Barakate A, Askari BM, Abbott JC, Ryan MD. Halpin C, et al. Plant Mol Biol. 2001 Sep;47(1-2):295-310. Plant Mol Biol. 2001. PMID: 11554478 Review.

References

1. 1. Nature. 1970 Aug 15;227(5259):680-5 - PubMed
2. 1. J Virol. 1982 Jan;41(1):244-9 - PubMed
3. 1. J Virol. 1985 Jun;54(3):651-60 - PubMed
4. 1. Virology. 1986 Feb;149(1):114-27 - PubMed
5. 1. Cell. 1986 Jun 6;45(5):761-70 - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Europe PubMed Central
  • Nature Publishing Group
  • PubMed Central
  • Wiley

• Other Literature Sources

  • The Lens - Patent Citations Database

• Miscellaneous

  • NCI CPTAC Assay Portal