Gemin5 proteolysis reveals a novel motif to identify L protease targets

David Piñeiro, Jorge Ramajo, Shelton Bradrick, Encarnación Martínez-Salas

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Translation of picornavirus RNA is governed by the internal ribosome entry site (IRES) element, directing the synthesis of a single polyprotein. Processing of the polyprotein is performed by viral proteases that also recognize as substrates host factors. Among these substrates are translation initiation factors and RNA-binding proteins whose cleavage is responsible for inactivation of cellular gene expression. Foot-and-mouth disease virus (FMDV) encodes two proteases, L pro and 3C pro. Widespread definition of L pro targets suffers from the lack of a sufficient number of characterized substrates. Here, we report the proteolysis of the IRES-binding protein Gemin5 in FMDV-infected cells, but not in cells infected by other picornaviruses. Proteolysis was specifically associated with expression of L pro, yielding two stable products, p85 and p57. In silico search of putative L targets within Gemin5 identified two sequences whose potential recognition was in agreement with proteolysis products observed in infected cells. Mutational analysis revealed a novel L pro target sequence that included the RKAR motif. Confirming this result, the Fas-ligand Daxx, was proteolysed in FMDV-infected and L pro-expressing cells. This protein carries a RRLR motif whose substitution to EELR abrogated L pro recognition. Thus, the sequence (R)(R/K)(L/A)(R) defines a novel motif to identify putative targets of L pro in host factors.

Original languageEnglish (US)
Pages (from-to)4942-4953
Number of pages12
JournalNucleic Acids Research
Volume40
Issue number11
DOIs
StatePublished - Jun 2012
Externally publishedYes

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Foot-and-Mouth Disease Virus
Proteolysis
Peptide Hydrolases
Picornaviridae
Polyproteins
Peptide Initiation Factors
Fas Ligand Protein
RNA-Binding Proteins
Computer Simulation
Carrier Proteins
RNA
Gene Expression
Proteins
Internal Ribosome Entry Sites

ASJC Scopus subject areas

  • Genetics

Cite this

Piñeiro, D., Ramajo, J., Bradrick, S., & Martínez-Salas, E. (2012). Gemin5 proteolysis reveals a novel motif to identify L protease targets. Nucleic Acids Research, 40(11), 4942-4953. https://doi.org/10.1093/nar/gks172

Gemin5 proteolysis reveals a novel motif to identify L protease targets. / Piñeiro, David; Ramajo, Jorge; Bradrick, Shelton; Martínez-Salas, Encarnación.

In: Nucleic Acids Research, Vol. 40, No. 11, 06.2012, p. 4942-4953.

Research output: Contribution to journalArticle

Piñeiro, D, Ramajo, J, Bradrick, S & Martínez-Salas, E 2012, 'Gemin5 proteolysis reveals a novel motif to identify L protease targets', Nucleic Acids Research, vol. 40, no. 11, pp. 4942-4953. https://doi.org/10.1093/nar/gks172
Piñeiro, David ; Ramajo, Jorge ; Bradrick, Shelton ; Martínez-Salas, Encarnación. / Gemin5 proteolysis reveals a novel motif to identify L protease targets. In: Nucleic Acids Research. 2012 ; Vol. 40, No. 11. pp. 4942-4953.
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