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Research article - Peer-reviewed, 2019

Structural characterization and biological function of bivalent binding of CD2AP to intrinsically disordered domain of chikungunya virus nsP3 protein

Agback, Peter; Dominguez, Francisco; Pustovalova, Yulia; Lukash, Tetyana; Shiliaev, Nikita; Orekhov, Vladislav Yu; Frolov, Ilya; Agback, Tatiana; Frolova, Elena I.


Alphavirus nsP3 proteins contain long, intrinsically disordered, hypervariable domains, HVD, which serve as hubs for interaction with many cellular proteins. Here, we have deciphered the mechanism and function of HVD interaction with host factors in alphavirus replication. Using NMR spectroscopy, we show that CHIKV HVD contains two SH3 domain-binding sites. Using an innovative chemical shift perturbation signature approach, we demonstrate that CD2AP interaction with HVD is mediated by its SH3-A and SH3-C domains, and this leaves the SH3-B domain available for interaction with other cellular factor(s). This cooperative interaction with two SH3 domains increases binding affinity to CD2AP and possibly induces long-range allosteric effects in HVD. Our data demonstrate that BIN1, CD2AP and SH3KBP1 play redundant roles in initiation of CHIKV replication. Point mutations in both CHIKV HVD binding sites abolish its interaction with all three proteins, CD2AP, BIN1 and SH3KBP1. This results in strong inhibition of viral replication initiation.


Alphaviruses; Chikungunya virus; Viral replication; Intrinsically disordered proteins; SH3 domain-containing proteins; CD2AP; SH3KBP1; BIN1; nsP3; NMR

Published in

2019, volume: 537, pages: 130-142

Authors' information

Swedish University of Agricultural Sciences, Department of Molecular Sciences
Dominguez, Francisco
University of Alabama at Birmingham (UAB)
Pustovalova, Yulia
University of Gothenburg
Lukash, Tetyana
University of Alabama at Birmingham (UAB)
Shiliaev, Nikita
University of Alabama at Birmingham (UAB)
Orekhov, Vladislav Yu
University of Gothenburg
Frolov, Ilya
University of Alabama at Birmingham (UAB)
Swedish University of Agricultural Sciences, Department of Molecular Sciences
Frolova, Elena I.
University of Alabama at Birmingham (UAB)

Sustainable Development Goals

SDG3 Good health and well-being

UKÄ Subject classification

Structural Biology
Biochemistry and Molecular Biology

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