The interdomain region of dengue NS5 protein that binds to the viral helicase NS3 contains independently functional importin β1 and importin α/β-recognized nuclear localization signals

Andrew J. Brooks, Magnus Johansson, Anna V. John, Yibin Xu, David A. Jans, Subhash G. Vasudevan

Research output: Contribution to journalArticleResearchpeer-review

118 Citations (Scopus)

Abstract

Dengue virus NS5 protein is a multifunctional RNA-dependent RNA polymerase that is essential for virus replication. We have shown previously that the 37-amino acid interdomain spacer sequence (residues 369 X 2 KKX 14 KKKX 11 RKX 3 405 ) of Dengue2 NS5 contains a functional nuclear localization signal (NLS). In this study, β-galactosidase fusion proteins carrying point mutations of the positively charged residues or truncations of the interdomain linker region (residues 369-389 or residues 386-405) were analyzed for nuclear import and importin binding activities to show that the N-terminal part of the linker region (residues 369-389, a/bNLS) is critical for nuclear localization and is recognized with high affinity by the conventional NLS-binding importin α/β heterodimeric nuclear import receptor. We also show that the importin β-binding site (residues 320-368, bNLS) adjacent to the a/bNLS, previously identified by yeast two-hybrid analysis, is functional as an NLS, recognized with high affinity by importin β, and able to target β-galactosidase to the nucleus. Intriguingly, the bNLS is highly conserved among Dengue and related flaviviruses, implying a general role for the region and importin β in the infectious cycle.

Original languageEnglish
Pages (from-to)36399-36407
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number39
DOIs
Publication statusPublished - 27 Sep 2002
Externally publishedYes

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