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Updated: Sep 20, 2025
RNA Interference in Ticks
Published on: January 20, 2011
Molecular basis of Tick Born encephalitis virus NS5 mediated subversion of apico-basal cell polarity signalling
Airah Javorsky1, Patrick O Humbert1,2,3,4, Marc Kvansakul1,2
1Department of Biochemistry & Chemistry, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria 3086, Australia.
Abstract:
The Scribble (Scrib) protein is a conserved cell polarity regulator with anti-tumorigenic properties. Viruses like the Tick-born encephalitis virus (TBEV) target Scribble to establish a cellular environment supporting viral replication, which is ultimately associated with poor prognosis upon infection. The TBEV NS5 protein has been reported to harbour both an internal as well as a C-terminal PDZ binding motif (PBM), however only the internal PBM was shown to be an interactor with Scribble, with the interaction being mediated via the Scribble PDZ4 domain to antagonize host interferon responses. We examined the NS5 PBM motif interactions with all Scribble PDZ domains using isothermal titration calorimetry, which revealed that the proposed internal PBM did not interact with any Scribble PDZ domains. Instead, the C-terminal PBM of NS5 interacted with Scrib PDZ3. We then established the structural basis of these interactions by determining crystal structures of Scrib PDZ3 bound to the NS5 C-terminal PBM. Our findings provide a structural basis for Scribble PDZ domain and TBEV NS5 interactions and provide a platform to dissect the pathogenesis of TBEV and the role of cell polarity signalling using structure guided approaches.
Insights
Tick-borne encephalitis virus (TBEV) NS5 protein interacts with Scribble PDZ3, not Scribble PDZ4, contrary to previous findings. This interaction, elucidated by structural analysis, offers insights into TBEV pathogenesis and cell polarity signaling.
Area of Science:
- Virology
- Molecular Biology
- Structural Biology
Background:
- Scribble (Scrib) protein regulates cell polarity and has anti-tumorigenic roles.
- Tick-borne encephalitis virus (TBEV) exploits Scribble to promote viral replication, correlating with poor patient prognosis.
- The TBEV NS5 protein possesses potential PDZ binding motifs (PBMs) implicated in Scribble interaction.
Purpose of the Study:
- To investigate the interaction between TBEV NS5 protein's PBMs and Scribble PDZ domains.
- To determine the structural basis of the TBEV NS5 and Scribble interaction.
- To provide a foundation for understanding TBEV pathogenesis and cell polarity signaling.
Main Methods:
- Isothermal titration calorimetry (ITC) to assess binding affinities.
- X-ray crystallography to determine the structure of interacting proteins.
- Analysis of PDZ domain interactions with viral protein motifs.
Main Results:
- The internal PBM of TBEV NS5 did not interact with any Scribble PDZ domains.
- The C-terminal PBM of TBEV NS5 specifically interacted with the Scribble PDZ3 domain.
- Crystal structures revealed the atomic details of the Scrib PDZ3-NS5 C-terminal PBM complex.
Conclusions:
- The interaction between TBEV NS5 and Scribble occurs via the C-terminal PBM binding to Scrib PDZ3.
- This study corrects previous assumptions about the NS5-Scribble interaction site.
- Structural insights pave the way for dissecting TBEV pathogenesis and cell polarity regulation.
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