Interaction of the intrinsically disordered C-terminal domain of the sesbania mosaic virus RNA-dependent RNA polymerase with the viral protein P10 in vitro: modulation of the oligomeric state and polymerase activity

Bakshi, Arindam and Sridhar, Shruthi and Sistla, Srinivas and Savithri, Handanahal Subbarao (2019) Interaction of the intrinsically disordered C-terminal domain of the sesbania mosaic virus RNA-dependent RNA polymerase with the viral protein P10 in vitro: modulation of the oligomeric state and polymerase activity. In: ARCHIVES OF VIROLOGY, 164 (4). pp. 971-982.

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Abstract

The RNA-dependent RNA polymerase (RdRp) of sesbania mosaic virus (SeMV) was previously shown to interact with the viral protein P10, which led to enhanced polymerase activity. In the present investigation, the equilibrium dissociation constant for the interaction between the two proteins was determined to be 0.09 mu M using surface plasmon resonance, and the disordered C-terminal domain of RdRp was shown to be essential for binding to P10. The association with P10 brought about a change in the oligomeric state of RdRp, resulting in reduced aggregation and increased polymerase activity. Interestingly, unlike the wild-type RdRp, C-terminal deletion mutants (C del 43 and C del 72) were found to exist predominantly as monomers and were as active as the RdRp-P10 complex. Thus, either the deletion of the C-terminal disordered domain or its masking by binding to P10 results in the activation of polymerase activity. Further, deletion of the C-terminal 85 residues of RdRp resulted in complete loss of activity. Mutation of a conserved tyrosine (RdRp Y480) within motif E, located between 72 and 85 residues from the C-terminus of RdRp, rendered the protein inactive, demonstrating the importance of motif E in RNA synthesis in vitro.

Item Type:	Journal Article
Publication:	ARCHIVES OF VIROLOGY
Publisher:	SPRINGER WIEN
Additional Information:	Copyright of this article belongs to SPRINGER WIEN
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	24 May 2019 12:23
Last Modified:	24 May 2019 12:23
URI:	http://eprints.iisc.ac.in/id/eprint/62362

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