University of Surrey

Test tubes in the lab Research in the ATI Dance Research

Structural and functional analysis of the Kaposi's sarcoma-associated herpesvirus vFLIP internal ribosome entry site.

Sulaiman, Mariam K. (2017) Structural and functional analysis of the Kaposi's sarcoma-associated herpesvirus vFLIP internal ribosome entry site. Doctoral thesis, University of Surrey.

Revised Thesis.pdf - Version of Record
Available under License Creative Commons Attribution Non-commercial Share Alike.

Download (5MB) | Preview


Kaposi's sarcoma-associated herpesvirus (KSHV) is an oncogenic virus, the etiological agent of Kaposi's sarcoma (KS) and primary effusion lymphoma (PEL). One of the key viral proteins that contribute to tumorigenesis is vFLIP, a viral homolog of the FLICE inhibitory protein. This KSHV protein interacts with the NFκB pathway to trigger the expression of antiapoptotic and proinflammatory genes and ultimately leads to tumor formation. The expression of vFLIP is regulated at the translational level by an internal ribosomal entry site (IRES) element. However, the precise mechanism by which ribosomes are recruited internally and the exact location of the IRES has remained elusive. The aims of this study were to confirm the previously identified 252-nt fragment directly upstream of vFLIP as the location of the vFLIP IRES in cellulo and to determine the structure and mechanism of action of the vFLIP IRES. Here we show that a 252-nt, within a coding region, directs translation in HEK293 cells. We have also established its RNA structure using chemical and enzymatic probing of RNA structure in solution and mutational analysis studies revealed that the domain If of the vFLIP IRES is crucial for its activity. Also, we demonstrate that IRES activity requires the presence of eIF4A and the eIF4E-eIF4G interaction. These interactions may define a new paradigm for IRES-mediated translation. Finally, we attempted to identify cellular proteins that may interact with the vFLIP IRES using several types of protein affinity chromatography, but we could detect a protein interacting with vFLIP IRES but yet to be confirmed.

Item Type: Thesis (Doctoral)
Divisions : Theses
Authors :
Sulaiman, Mariam K.
Date : 31 October 2017
Contributors :
Depositing User : Mariam Sulaiman
Date Deposited : 10 Nov 2017 10:58
Last Modified : 10 Nov 2017 10:58

Actions (login required)

View Item View Item


Downloads per month over past year

Information about this web site

© The University of Surrey, Guildford, Surrey, GU2 7XH, United Kingdom.
+44 (0)1483 300800