We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Repression of the Internal Ribosome Entry Site-dependent Translation of Hepatitis C Virus by an Engineered PUF Protein.
- Authors
Kiani, Seyed Jalal; Taheri, Tahereh; Nejati, Ahmad; Maleki, Monireh; Rafati, Sima; Azadmanesh, Kayhan; Alavian, Seyed Moayed; Azad, Talat Mokhtari; Samimi-Rad, Katayoun
- Abstract
Background: Pumilio/fem-3mRNA binding factor (PUF) proteins can bind RNA in a sequence-specific manner. The deciphered RNA-recognition code of these proteins has enabled researchers to design engineered PUF proteins, capable of binding to any desired target in order to modify its ultimate fate. In this study, a modified Homo sapiens Pumilio 1-homology domain (HsPUM1-HD) was engineered to bind to the internal ribosome entry site (IRES) of hepatitis C virus (HCV) genome to potentially inhibit viral translation. Methods: Based on the RNA-recognition code, required modifications were applied to HsPUM1-HD in order to change its natural recognition sequence to a sequence in the stem-loop III of HCV IRES. RNA protein pull-down assay was performed to assess the sequence specificity of the modified HsPUM1-HD (mHsPUM1-HD). Translational inhibitory effect of mHsPUM1-HD was evaluated in a dual-luciferase reporter assay. Results: The mHsPUM1-HD was found to bind to its cognate RNA in a sequence-specific manner, as a biotinylated target RNA captured mHsPUM1-HD through binding to streptavidin magnetic beads. This protein also reduced HCV IRES-dependent firefly luciferase translation by 40% in HEK293 cells. Conclusions: The present study is the first report of an engineered HsPUM1-HD with potential anti-HCV activity. These findings suggest that PUM-HDs can be engineered to target desired RNAs of infectious agents in order to specifically interrupt protein translation, as an essential step of their life cycle.
- Subjects
ANALYSIS of variance; GENETIC engineering; HEPATITIS C; HEPATITIS viruses; HUMAN genome; WESTERN immunoblotting; DATA analysis software; DESCRIPTIVE statistics; SEQUENCE analysis; GENOTYPES; ONE-way analysis of variance; RNA-binding proteins
- Publication
Hepatitis Monthly, 2017, Vol 17, Issue 2, p1
- ISSN
1735-143X
- Publication type
Article
- DOI
10.5812/hepatmon.45022