Title
Functional analysis of the interaction between HCV 5'UTR and putative subunits of eukaryotic translation initiation factor eIF3.
Abstract
Translation initiation in Hepatitis C Virus is controlled by the presence of an internal ribosome entry site element (IRES) principally located in its 5' untranslated region (UTR). Mutation/deletion analyses have shown that the integrity of this structure is essential for initiation of cap-independent protein synthesis. We have developed a strategy to swap the position of the two-major domains (II and III) on the 5'UTR sequence. The aim was to further characterize this mechanism by preserving domain-specific interactions but possibly losing contacts that require any interdomain geometry. The expression of dicistronic mRNAs containing these different UTRs showed that the positioning of the different domains on the 5'UTR is essential for efficient IRES functioning. We then used these mutants to identify cellular factors implicated in IRES activity. Using UV crosslinking assays we found that domain III makes direct contact with two proteins (p170/p120) which can be associated with efficient IRES activity. In particular, we have mapped the binding sites of these proteins and shown that p120 binds to the apical loop segment of domain III, whilst p170 binds in the stem portion, independently of domain III position or context. Finally, we provide evidence showing that p170 and p120 represent two subunits of eukaryotic initiation factor eIF3: p170 and p116/p110.
Year
DOI
Venue
1998
10.1093/nar/26.13.3179
NUCLEIC ACIDS RESEARCH
Keywords
Field
DocType
functional analysis,cos cells,protein binding,rna binding proteins,internal ribosome entry site,translation initiation,eukaryotic initiation factor,untranslated region,binding site,protein synthesis
Untranslated region,Biology,Hepatitis C virus internal ribosome entry site,Internal ribosome entry site,Initiation factor,Eukaryotic initiation factor,Five prime untranslated region,Eukaryotic translation,Eukaryotic translation initiation factor 4 gamma,Genetics,Molecular biology
Journal
Volume
Issue
ISSN
26
13.0
0305-1048
Citations 
PageRank 
References 
0
0.34
0
Authors
4
Name
Order
Citations
PageRank
Emanuele Buratti110.71
S Tisminetzky200.34
M Zotti300.34
F E Baralle401.01