Journal
PROTEOMICS
Volume 8, Issue 22, Pages 4782-4790Publisher
WILEY
DOI: 10.1002/pmic.200800338
Keywords
Internal initiation of translation; Hepatitis C; Mass spectrometry; Picornavirus; RNA-binding proteins
Funding
- Comunidad Autonoma de Madrid [BFU2005-00948]
- Fundacion Ramon Areces
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Initiation of translation driven by internal ribosome entry site (IRES) elements depends upon the structural organization of this mRNA region. Besides translation initiation factors (eIFs), auxiliary proteins can also affect I RES activity. With the aim to identify proteins interacting with two unrelated IRESs present in the genome of foot-and-mouth disease virus (FM DV) and hepatitis C virus (HCV) we have used a proteomic approach. This procedure allowed the identification of 21 RNA-binding proteins interacting with discrete regions of the FMDV IRES, domains 3 and 5, and 16 interacting with domain I IT of the HCV IRES. In support of the binding specificity the factors interacting with domain 3 differed from those interacting with domain 5, and included three poly(rC)-binding protein (PCBP) members, besides proliferation-associated 2G4 (PA2G4) and deleted-azoospermia 1 (DAZ1) protein. Around 71% of the identified factors associated with the FMDV IRES differ from those interacting with the HCV IRES. The group of proteins interacting with the FMDV or the HCV TRES includes eIF4B and 5 subunits of eIF3, respectively, known to interact with each of these RNAs, validating the results of this approach. According to the function of the identified proteins, 55% are involved in translation control, whereas 35% play a role in different aspects of RNA lifespan. Compilation of factors preferentially associated with FMDV or HCV TRES provides a basis for examining the strategies used by IRESs to recruit the translation machinery.
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