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| Comparison of Iron Responsive Element (IRE) and the Hepatitis C Virus (HCV) Internal Ribosome Entry Site (IRES)-Domain IV: Structure and Function |
| Category:
Flaviviridae - Other |
Ebenezer Tumban, Jenna Painta, William B. Lott
New Mexico State University, Las Cruces, NM |
| Presentation Number: 267 |
| Keyword:
HCV IRES, IRE, Gel mobility shift assay |
| Some eukaryotic mRNAs contain stem-loop structures at their 5' and 3' untranslated regions (UTRs) known as iron responsive element (IRE) that are used in regulating the translation of host proteins. A structure very similar to the eukaryotic IRE, containing a base-paired stem loop , a terminal loop, and a putative G-C base-pair in the loop is found in Domain IV, a region of unknown function, of the hepatitis C virus (HCV) internal ribosomal entry site (IRES). In the current study we investigated whether HCV possesses an element in its IRES that is or can act as an IRE by binding domain IV to human iron regulatory proteins (hIRP1s). We used an oligonucleotide generated mutagenesis approach to generate seven mutants (deletions and substitution mutations) of Domain IV. These mutants have been tested with hIRP1 in a gel mobility shift assay. Results from this study show that Domain IV is not an authentic IRE but some of its mutants (ET3, ET1+3, ET2+3, and ET1+2+3) could bind to hIRP1. The Kd value for mutant ET2+3 with hIRP1 was determined in this study and is very high compared to a consensus IRE, showing a low binding affinity. Although domain IV is not an authentic IRE, we believe that hIRP1 can be retargeted to bind domain IV by mutating some amino acids in the binding domain of hIRP1. This approach shows promise for producing a lead compound for HCV-specific antiviral drug design targeting the mechanism of viral translation. |
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