Analysis of self-association of West Nile virus capsid protein and the crucial role played by Trp 69 in homodimerization |
| |
Authors: | Bhuvanakantham Raghavan Ng Mah-Lee |
| |
Institution: | Flavivirology Laboratory, Department of Microbiology, 5 Science Drive 2, National University of Singapore, Singapore 117597, Singapore. |
| |
Abstract: | The understanding of capsid (C) protein interactions with itself would provide important data on how the core is organized in flaviviruses during assembly. In this study, West Nile (WN) virus C protein was shown to form homodimers using yeast two-hybrid analysis in conjunction with mammalian two-hybrid and in vivo co-immunoprecipitation assays. To delineate the region on the C protein which mediates C-C dimerization, truncation studies were carried out. The results obtained clearly showed that the internal hydrophobic segment flanked by helix I and helix III of WN virus C protein is essential for the self-association of C protein. The crucial role played by Trp 69 in stabilizing the self-association of C protein was also demonstrated by mutating Trp to Gly/Arg/Phe. Substitution of the Trp residue with Gly/Arg abolished the dimerization, whereas substitution with Phe decreased the self-association significantly. The results of this study pinpoint a critical residue in the C protein that potentially plays a role in stabilizing the homotypic interaction. |
| |
Keywords: | Capsid Tryptophan Homotypic interaction Yeast two-hybrid Mammalian two-hybrid Co-immunoprecipitation |
本文献已被 ScienceDirect PubMed 等数据库收录! |